Inhibitors of akt activity

ABSTRACT

The present invention is directed to compounds which contain substituted pyridazines and pyrimidines moieties which inhibit the activity of Akt, a serine/threonine protein kinase. The invention is further directed to chemotherapeutic compositions containing the compounds of this invention and methods for treating cancer comprising administration of the compounds of the invention.

BACKGROUND OF THE INVENTION

The present invention relates to compounds which contain substitutedpyridazines and pyrimidines that are inhibitors of the activity of oneor more of the isoforms of the serine/threonine kinase, Akt (also knownas PKB; hereinafter referred to as “Akt”). The present invention alsorelates to pharmaceutical compositions comprising such compounds andmethods of using the instant compounds in the treatment of cancer.

Apoptosis (programmed cell death) plays essential roles in embryonicdevelopment and pathogenesis of various diseases, such as degenerativeneuronal diseases, cardiovascular diseases and cancer. Recent work hasled to the identification of various pro- and anti-apoptotic geneproducts that are involved in the regulation or execution of programmedcell death. Expression of anti-apoptotic genes, such as Bcl2 orBc1-x_(L), inhibits apoptotic cell death induced by various stimuli. Onthe other hand, expression of pro-apoptotic genes, such as Bax or Bad,leads to programmed cell death (Adams et al. Science, 281:1322-1326(1998)). The execution of programmed cell death is mediated by caspase-1related proteinases, including caspase-3, caspase-7, caspase-8 andcaspase-9 etc (Thornberry et al. Science, 281:1312-1316 (1998)).

The phosphatidylinositol 3′-OH kinase (PI3K)/Akt pathway appearsimportant for regulating cell survival/cell death (Kulik et al. Mol.Cell. Biol. 17:1595-1606 (1997); Franke et al, Cell, 88:435-437 (1997);Kauffmann-Zeh et al. Nature 385:544-548 (1997) Hemmings Science,275:628-630 (1997); Dudek et al., Science, 275:661-665 (1997)). Survivalfactors, such as platelet derived growth factor (PDGF), nerve growthfactor (NGF) and insulin-like growth factor-1 (IGF-1), promote cellsurvival under various conditions by inducing the activity of PI3K(Kulik et al. 1997, Hemmings 1997). Activated PI3K leads to theproduction of phosphatidylinositol (3,4,5)-triphosphate(PtdIns(3,4,5)-P3), which in turn binds to, and promotes the activationof, the serine/threonine kinase Akt, which contains a pleckstrinhomology (PH)-domain (Franke et al Cell, 81:727-736 (1995); HemmingsScience, 277:534 (1997); Downward, Curr. Opin. Cell Biol. 10:262-267(1998), Alessi et al., EMBO J. 15: 6541-6551 (1996)). Specificinhibitors of PI3K or dominant negative Akt mutants abolishsurvival-promoting activities of these growth factors or cytokines. Ithas been previously disclosed that inhibitors of PI3K (LY294002 orwortmannin) blocked the activation of Akt by upstream kinases. Inaddition, introduction of constitutively active PI3K or Akt mutantspromotes cell survival under conditions in which cells normally undergoapoptotic cell death (Kulik et al. 1997, Dudek et al. 1997).

Three members of the Akt subfamily of second-messenger regulatedserine/threonine protein kinases have been identified and termedAkt1/PKBα, Akt2/PKBβ, and Akt3/PKBγ (hereinafter referred to as “Akt1”,“Akt2” and “Akt3”), respectively. The isoforms are homologous,particularly in regions encoding the catalytic domains. Akts areactivated by phosphorylation events occurring in response to PI3Ksignaling. PI3K phosphorylates membrane inositol phospholipids,generating the second messengers phosphatidyl-inositol3,4,5-trisphosphate and phosphatidylinositol 3,4-bisphosphate, whichhave been shown to bind to the PH domain of Akt. The current model ofAkt activation proposes recruitment of the enzyme to the membrane by3′-phosphorylated phosphoinositides, where phosphorylation of theregulatory sites of Akt by the upstream kinases occurs (B. A. Hemmings,Science 275:628-630 (1997); B. A. Hemmings, Science 276:534 (1997); J.Downward, Science 279:673-674 (1998)).

Phosphorylation of Aka occurs on two regulatory sites, Thr³⁰⁸ in thecatalytic domain activation loop and on Ser⁴⁷³ near the carboxy terminus(D. R. Alessi et al. EMBO J. 15:6541-6551 (1996) and R. Meier et al. J.Biol. Chem. 272:30491-30497 (1997)). Equivalent regulatoryphosphorylation sites occur in Akt2 and Akt3. The upstream kinase, whichphosphorylates Akt at the activation loop site has been cloned andtermed 3′-phosphoinositide dependent protein kinase 1 (PDK1). PDK1phosphorylates not only Akt, but also p70 ribosomal S6 kinase, p90RSK,serum and glucocorticoid-regulated kinase (SGK), and protein kinase C.The upstream kinase phosphorylating the regulatory site of Akt near thecarboxy terminus has not been identified yet, but recent reports imply arole for the integrin-linked kinase (ILK-1), a serine/threonine proteinkinase, or autophosphorylation.

Analysis of Akt levels in human tumors showed that Akt2 is overexpressedin a significant number of ovarian (J. Q. Cheng et al. Proc. Natl. Acad.Sci. U.S.A. 89:9267-9271 (1992)) and pancreatic cancers (J. Q. Cheng etal. Proc. Natl. Acad. Sci. U.S.A. 93:3636-3641 (1996)). Similarly, Akt3was found to be overexpressed in breast and prostate cancer cell lines(Nakatani et al. J. Biol. Chem. 274:21528-21532 (1999).

The tumor suppressor PTEN, a protein and lipid phosphatase thatspecifically removes the 3′ phosphate of PtdIns(3,4,5)-P3, is a negativeregulator of the PI3K/Akt pathway (Li et al. Science 275:1943-1947(1997), Stambolic et al. Cell 95:29-39 (1998), Sun et al. Proc. Natl.Acad. Sci. U.S.A. 96:6199-6204 (1999)). Germline mutations of PTEN areresponsible for human cancer syndromes such as Cowden disease (Liaw etal. Nature Genetics 16:64-67 (1997)). PTEN is deleted in a largepercentage of human tumors and tumor cell lines without functional PTENshow elevated levels of activated Akt (Li et al. supra, Guldberg et al.Cancer Research 57:3660-3663 (1997), Risinger et al. Cancer Research57:4736-4738 (1997)).

These observations demonstrate that the PI3K/Akt pathway plays importantroles for regulating cell survival or apoptosis in tumorigenesis.

Inhibition of Akt activation and activity can be achieved by inhibitingPI3K with inhibitors such as LY294002 and wortmannin. However, PI3Kinhibition has the potential to indiscriminately affect not just allthree Akt isozymes but also other PH domain-containing signalingmolecules that are dependent on PdtIns(3,4,5)-P3, such as the Tec familyof tyrosine kinases. Furthermore, it has been disclosed that Akt can beactivated by growth signals that are independent of PI3K.

Alternatively, Akt activity can be inhibited by blocking the activity ofthe upstream kinase PDK1. No specific PDK1 inhibitors have beendisclosed. Again, inhibition of PDK1 would result in inhibition ofmultiple protein kinases whose activities depend on PDK1, such asatypical PKC isoforms, SGK, and S6 kinases (Williams et al. Curr. Biol.10:439-448 (2000).

It is an object of the instant invention to provide novel compounds thatare inhibitors of Akt.

It is also an object of the present invention to provide pharmaceuticalcompositions that comprise the novel compounds that are inhibitors ofAkt.

It is also an object of the present invention to provide a method fortreating cancer that comprises administering such inhibitors of Aktactivity.

SUMMARY OF THE INVENTION

The instant invention provides for compounds which comprise substitutedpyridazines and pyrimidines that inhibit Akt activity. In particular,the compounds disclosed selectively inhibit one or two of the Aktisoforms. The invention also provides for compositions comprising suchinhibitory compounds and methods of inhibiting Akt activity byadministering the compound to a patient in need of treatment of cancer.

DETAILED DESCRIPTION OF THE INVENTION

The compounds of the instant invention are useful in the inhibition ofthe activity of the serine/threonine kinase Akt. In a first embodimentof this invention, the inhibitors of Akt activity are illustrated by theFormula A:

wherein:

X is N and Y is CH or X is CH and Y is N;

Q is: heterocyclyl, which is optionally substituted with 1-3 R^(z);

a is 0 or 1; b is 0 or 1; m is 0, 1 or 2; n is 0, 1 or 2; r is 0 or 1; sis 0 or 1;

R¹ is independently selected from: (C═O)_(a)O_(b)C₁-C₁₀ alkyl,(C═O)_(a)O_(b)aryl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, (C═O)_(a)O_(b)heterocyclyl, (C═O)_(a)O_(b)C₃-C₈ cycloalkyl, CO₂H, halo, CN, OH,O_(b)C₁-C₆ perfluoroalkyl, O_(a)(C═O)_(b)NR²R³, NR^(c)(C═O)_(b)NR²R³,S(O)_(m)R^(a), S(O)₂NR²R³, NR^(c)S(O)_(m)R^(a), oxo, CHO, NO₂,NR^(c)(C═O)O_(b)R^(a), O(C═O)O_(b)C₁-C₁₀ alkyl, O(C═O)O_(b)C₃-C₈cycloalkyl, O(C═O)O_(b)aryl, C₁-C₆alkyl(C═NR^(b))N(R^(b))₂;O(C═O)O_(b)-heterocycle, and O_(a)-P═O(OH)₂, said alkyl, aryl, alkenyl,alkynyl, heterocyclyl, and cycloalkyl optionally substituted with one ormore substituents selected from R^(z);

R² and R³ are independently selected from: H, (C═O)O_(b)R^(a), C₁-C₁₀alkyl, aryl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, heterocyclyl, C₃-C₉cycloalkyl, SO₂R^(a), (C═O)NR^(b) ₂, OH, and O_(a)-P═O(OH)₂, said alkyl,cycloalkyl, aryl, heterocylyl, alkenyl, and alkynyl is optionallysubstituted with one or more substituents selected from R^(z), or R² andR³ can be taken together with the nitrogen to which they are attached toform a monocyclic or bicyclic heterocycle with 4-7 members in each ringand optionally containing, in addition to the nitrogen, one or moreadditional heteroatoms selected from N, O and S, said monocyclic orbicyclic heterocycle optionally substituted with one or moresubstituents selected from R^(z);

R^(z) is selected from: (C═O)_(r)O_(s)(C₁-C₁₀)alkyl,O_(r)(C₁-C₃)perfluoroalkyl, (C₀-C₆)alkylene-S(O)_(m)R^(a), oxo, OH,halo, CN, (C═O)_(r)O_(s)(C₂-C₁₀)alkenyl, (C═O)_(r)O_(s)(C₂-C₁₀)alkynyl,(C═O)_(r)O_(s)(C₃-C₆)cycloalkyl, (C═O)_(r)O_(s)(C₀-C₆)alkylene-aryl,(C═O)_(r)O_(s)(C₀-C₆)alkylene-heterocyclyl,(C═O)_(r)O_(s)(C₀-C₆)alkylene-N(R^(b))₂, C(O)R^(a),(C₀-C₆)alkylene-CO₂R^(a), C(O)H, (C₀-C₆)alkylene-CO₂H, C(O)N(R^(b))₂,S(O)_(m)R^(a), S(O)₂N(R^(b))₂NR^(c)(C═O)O_(b)R^(a),O(C═O)O_(b)C₁-C₁₀alkyl, O(C═O)O_(b)C₃-C₈ cycloalkyl, O(C═O)O_(b)aryl,O(C═O)O_(b)-heterocycle, and O_(a)-P═O(OH)₂, said alkyl, alkenyl,alkynyl, cycloalkyl, aryl, and heterocyclyl is optionally substitutedwith up to three substituents selected from R^(b), OH, (C₁-C₆)alkoxy,halogen, CO₂H, CN, O(C═O)C₁-C₆ alkyl, oxo, N(R^(b))₂ and O_(a)-P═O(OH)₂;

R^(a) is: substituted or unsubstituted (C₁-C₆)alkyl, substituted orunsubstituted (C₂-C₆)alkenyl, substituted or unsubstituted(C₂-C₆)alkynyl, substituted or unsubstituted (C₃-C₆)cycloalkyl,substituted or unsubstituted aryl, (C₁-C₆)perfluoroalkyl,2,2,2-trifluoroethyl, or substituted or unsubstituted heterocyclyl;

R^(b) is independently: H, (C₁-C₆)alkyl, substituted or unsubstitutedaryl, substituted or unsubstituted benzyl, substituted or unsubstitutedheterocyclyl, (C₃-C₆)cycloalkyl, (C═O)OC₁-C₆ alkyl, (C═O)C₁-C₆ alkyl orS(O)₂R^(a); and

R^(c) is selected from: H, C₁-C₁₀ alkyl, aryl, C₂-C₁₀ alkenyl, C₂-C₁₀alkynyl, heterocyclyl, C₃-C₉ cycloalkyl, and C₁-C₆ perfluoroalkyl, saidalkyl, cycloalkyl, aryl, heterocylyl, alkenyl, and alkynyl is optionallysubstituted with one or more substituents selected from R^(z); or apharmaceutically acceptable salt or a stereoisomer thereof

In a second embodiment of the instant invention is a compoundillustrated by Formula B-1:

wherein:

Q is: heterocyclyl, which is optionally substituted with one to threeR^(z);

R¹ is selected from: NH₂; halogen; OH; oxo; CN; phenyl; heterocyclyl;(C═O)_(r)O_(s)(C₀-C₆)alkyl; (C₀-C₆)alkyl-NR^(a)R^(b); andO_(a)-P═O(OH)₂; wherein R^(a) and R^(b) are independently H or(C₁-C₆)alkyl and wherein said alkyl and heterocyclyl are optionallysubstituted with OH, oxo, O_(a)-P═O(OH)₂ and O(C₁-C₆)alkyl; and allother substituents and variables are as defined in the first embodiment;or a pharmaceutically acceptable salt or a stereoisomer thereof.

In a third embodiment of the instant invention is a compound illustratedby Formula B-2:

wherein:

all substituents and variables are as defined in the second embodiment;or a pharmaceutically acceptable salt or a stereoisomer thereof.

Specific compounds of the instant invention include:

-   N,N-dimethyl-5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-pyrazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazin-3-amine    (1-5);-   N-methyl-5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-pyrazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazin-3-amine    (1-6);-   4-[5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)-piperidin-1-yl]methyl}phenyl)pyridazin-3-yl]morpholine    (1-7);-   tert-butyl    4-[5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazin-3-yl]piperazine-1-carboxylate    (1-8);-   4-phenyl-6-piperazin-1-yl-3-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazine    (1-9);-   4-[5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazin-3-yl]thiomorpholine    (1-10);-   4-[5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazin-3-yl]thiomorpholine-1-oxide    (1-11);-   4-[5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazin-3-yl]thiomorpholine-1,1-dioxide    (1-12);-   3-methoxy-5-phenyl-6-(4-{[4-(5pyridin-2-yl-1H-pyrazol-3-yl)piperidin-1-yl]methyl}-phenyl)pyridazine    (2-3);-   5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-pyrazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazin-3-ol    (2-4);-   4-phenyl-6-pyridin-3-yl-3-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazine    (3-4);-   5-phenyl-2-pyridin-2-yl-4-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidine    (4-4); and-   5-phenyl-4-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidin-2-amine    (4-5); or a pharmaceutically acceptable salt or a stereoisomer    thereof.

Specific TFA salts of the compounds of the instant invention include:

-   N-methyl-5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-pyrazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazin-3-amine    (1-6);-   3-methoxy-5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-pyrazol-3-yl)piperidin-1-yl]methyl}-phenyl)pyridazine    (2-3); and-   5-phenyl-2-pyridin-2-yl-4-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidine    (4-4); or a stereoisomer thereof.

In a further embodiment, a specific compound of the instant inventionis:

-   N,N-dimethyl-5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-pyrazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazin-3-amine    (1-5); and-   5-phenyl-2-pyridin-2-yl-4-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidine    (4-4); or a pharmaceutically acceptable salt or a stereoisomer    thereof.

In another embodiment, compounds of the instant invention include:

-   N,N-dimethyl-5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-pyrazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazin-3-amine    (1-5);-   N-methyl-5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-pyrazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazin-3-amine    (1-6);-   4-[5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)-piperidin-1-yl]methyl}phenyl)pyridazin-3-yl]morpholine    (1-7);-   tert-butyl    4-[5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazin-3-yl]piperazine-1-carboxylate    (1-8);-   4-phenyl-6-piperazin-1-yl-3-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazine    (1-9);-   4-[5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazin-3-yl]thiomorpholine    (1-10);-   4-[5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazin-3-yl]thiomorpholine-1-oxide    (1-11);-   4-[5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazin-3-yl]thiomorpholine-1,1-dioxide    (1-12);-   3-methoxy-5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-pyrazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazine    (2-3);-   5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-pyrazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazin-3-ol    (2-4);-   4-phenyl-6-pyridin-3-yl-3-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazine    (3-4);-   5-phenyl-2-pyridin-2-yl-4-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidine    (4-4);-   5-phenyl-4-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidin-2-amine    (4-5);-   1-{-4-[5-phenyl-2-(pyridine-3-ylamino)pyrimidin-4-yl]benzyl}-4-(5-pyridin-2-yl-4H-1,2,4-triazol-3-yl)piperidine    (5-6);-   1-{-4-[2-(5-amino-1,3,4-thiadiazol-2-yl)-5-phenylpyrimidin-4-yl]benzyl}-4-(5-pyridin-2-yl-4H-1,2,4-triazol-3-yl)piperidine    (5-7);-   1-methyl-4-(2-{[5-phenyl-4-(4-{[4-(5-pyridin-2-yl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidin-2-yl]amino}ethyl)piperzine    (5-8);-   1-[4-(2-amino-5-phenylpyrimidin-4-yl)benzyl]-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine    (5-9);-   1-{4-[2-(methylthio)-5-phenylpyrimidin-4-yl]benzyl}-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine    (5-10);-   1-{4-[2-(4-acetylpiperazin-1-O-5-phenylpyrimidin-4-yl]benzyl}-4-(5-pyridin-2-yl-4-h-1,2,4-triazol-3-yl)piperidine    (5-11);-   1-[4-(2-{[1-(ethoxycarbonyl)piperidin-4-yl]amino}-5-phenylpyrimidin-4-yl)benzyl]-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine    (5-12);-   1-methyl-4-[5-phenyl-4-(4-{[4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidin-2-yl]piperazine    (5-13);-   1-(2-hydroxyethyl)-4-[5-phenyl-4-(4-{[4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidin-2-yl]piperazine    (5-14);-   1-[2-(dimethylamino)ethyl]-4-[5-phenyl-4-(4-{[4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidin-2-yl]piperazine    (5-15);-   1-(4-{5-phenyl-2-[(pyridin-3-ylmethyl)amino]pyrimidin-4-yl}benzyl)-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine    (5-16);-   2-{[5-phenyl-4-(4-{[4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidin-2-yl]amino}pyridine    (5-17);-   2-{5-[1-(4-{2-[methyl(pyridin-2-yl)amino]-5-phenylpyrimidin-4-yl}benzyl)piperidin-4-yl]-4h-1,2,4-triazol-3-yl}pyridine    (5-18);-   1-{4-[2-(dimethylamino)-5-phenylpyrimidin-4-yl]benzyl}-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine    (5-19);-   1-(4-{2-[[2-(dimethylamino)ethyl](methyl)amino]-5-phenylpyrimidin-4-yl}benzyl)-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine    (5-20);-   1-[4-(2-{[3-(dimethylammonio)propyl]amino}-5-phenylpyrimidin-4-yl)benzyl]-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine    (5-21);-   1-[4-(2-{[2-(dimethylammonio)ethyl]amino}-5-phenylpyrimidin-4-yl)benzyl]-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine    (5-22); and-   1-(4-{2-[[3-(dimethylammonio)propyl](methyl)amino]-5-phenylpyrimidin-4-yl}benzyl)-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine    (5-23);    or a pharmaceutically acceptable salt or a stereoisomer thereof.

In another embodiment, specific TFA salts of the compounds of theinstant invention include:

-   N-methyl-5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-pyrazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazin-3-amine    (1-6);-   3-methoxy-5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-pyrazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazine    (2-3);-   5-phenyl-2-pyridin-2-yl-4-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidine    (4-4);

1-{4-[5-phenyl-2-(pyridine-3-ylamino)pyrimidin-4-yl]benzyl}-4-(5-pyridin-2-yl-4H-1,2,4-triazol-3-yl)piperidine(5-6);

-   1-{4-[2-(5-amino-1,3,4-thiadiazol-2-yl)-5-phenylpyrimidin-4-yl]benzyl}-4-(5-pyridin-2-yl-4H-1,2,4-triazol-3-yl)piperidine    (5-7);-   1-methyl-4-(2-{[5-phenyl-4-(4-{[4-(5-pyridin-2-yl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidin-2-yl]amino}ethyl)piperzine    (5-8);-   1-[4-(2-amino-5-phenylpyrimidin-4-yl)benzyl]-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine    (5-9);-   1-{4-[2-(methylthio)-5-phenylpyrimidin-4-yl]benzyl}-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine    (5-10);-   1-{4-[2-(4-acetylpiperazin-1-O-5-phenylpyrimidin-4-yl]benzyl}-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine    (5-11);-   1-[4-(2-{[1-(ethoxycarbonyl)piperidin-4-yl]amino}-5-phenylpyrimidin-4-yl)benzyl]-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine    (5-12);-   1-methyl-4-[5-phenyl-4-(4-{[4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidin-2-yl]piperazine    (5-13);-   1-(2-hydroxyethyl)-4-[5-phenyl-4-(4-{[4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidin-2-yl]piperazine    (5-14);-   1-[2-(dimethylamino)ethyl]-4-[5-phenyl-4-(4-{[4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidin-2-yl]piperazine    (5-15);-   1-(4-{5-phenyl-2-[(pyridin-3-ylmethyl)amino]pyrimidin-4-yl}benzyl)-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine    (5-16);-   2-{[5-phenyl-4-(4-{[4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidin-2-yl]amino}pyridine    (5-17);-   2-{5-[1-(4-{[4-methyl(pyridin-2-yl)amino]-5-phenylpyrimidin-4-yl}benzyl)piperidin-4-yl]-4h-1,2,4-triazol-3-yl}pyridine    (5-18);-   1-{4-[2-(dimethylamino)-5-phenylpyrimidin-4-yl]benzyl}-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine    (5-19);-   1-(4-{2-[[2-(dimethylamino)ethyl](methyl)amino]-5-phenylpyrimidin-4-yl}benzyl)-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine    (5-20);-   1-[4-(2-{[3-(dimethylammonio)propyl]amino}-5-phenylpyrimidin-4-yl)benzyl]-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine    (5-21);-   1-[4-(2-{[2-(dimethylammonio)ethyl]amino}-5-phenylpyrimidin-4-yl)benzyl]-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine    (5-22); and-   1-(4-{2-[[3-(dimethylammonio)propyl](methyl)amino]-5-phenylpyrimidin-4-yl}benzyl)-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine    (5-23);    or a stereoisomer thereof

The compounds of the present invention may have asymmetric centers,chiral axes, and chiral planes (as described in: E. L. Eliel and S. H.Wilen, Stereochemistry of Carbon Compounds, John Wiley & Sons, New York,1994, pages 1119-1190), and occur as racemates, racemic mixtures, and asindividual diastereomers, with all possible isomers and mixturesthereof, including optical isomers, all such stereoisomers beingincluded in the present invention.

In addition, the compounds disclosed herein may exist as tautomers andboth tautomeric forms are intended to be encompassed by the scope of theinvention, even though only one tautomeric structure is depicted. Forexample, any claim to compound A below is understood to includetautomeric structure B, and vice versa, as well as mixtures thereof. Thetwo tautomeric forms of the benzimidazolonyl moiety are also within thescope of the instant invention.

Tetrazoles exist as a mixture of 1H/2H tautomers. The tautomeric formsof the tetrazol moiety are also within the scope of the instantinvention.

When any variable (e.g. R¹, R^(z), etc.) occurs more than one time inany constituent, its definition on each occurrence is independent atevery other occurrence. Also, combinations of substituents and variablesare permissible only if such combinations result in stable compounds.Lines drawn into the ring systems from substituents represent that theindicated bond may be attached to any of the substitutable ring atoms.If the ring system is polycyclic, it is intended that the bond beattached to any of the suitable carbon atoms on the proximal ring only.

It is understood that substituents and substitution patterns on thecompounds of the instant invention can be selected by one of ordinaryskill in the art to provide compounds that are chemically stable andthat can be readily synthesized by techniques known in the art, as wellas those methods set forth below, from readily available startingmaterials. If a substituent is itself substituted with more than onegroup, it is understood that these multiple groups may be on the samecarbon or on different carbons, so long as a stable structure results.The phrase “optionally substituted with one or more substituents” shouldbe taken to be equivalent to the phrase “optionally substituted with atleast one substituent” and in such cases an embodiment will have fromzero to four substituents, and another embodiment will have from zero tothree substituents.

As used herein, “alkyl” is intended to include both branched andstraight-chain saturated aliphatic hydrocarbon groups having thespecified number of carbon atoms. For example, C₁-C₁₀, as in “C₁-C₁₀alkyl” is defined to include groups having 1, 2, 3, 4, 5, 6, 7, 8, 9 or10 carbons in a linear or branched arrangement. For example, “C₁-C₁₀alkyl” specifically includes methyl, ethyl, n-propyl, i-propyl, n-butyl,t-butyl, i-butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, and so on.The term “cycloalkyl” means a monocyclic saturated aliphatic hydrocarbongroup having the specified number of carbon atoms. For example,“cycloalkyl” includes cyclopropyl, methyl-cyclopropyl,2,2-dimethyl-cyclobutyl, 2-ethyl-cyclopentyl, cyclohexyl, and so on.

“Alkoxy” represents either a cyclic or non-cyclic alkyl group ofindicated number of carbon atoms attached through an oxygen bridge.“Alkoxy” therefore encompasses the definitions of alkyl and cycloalkylabove.

If no number of carbon atoms is specified, the term “alkenyl” refers toa non-aromatic hydrocarbon radical, straight, branched or cyclic,containing from 2 to 10 carbon atoms and at least one carbon to carbondouble bond. Preferably one carbon to carbon double bond is present, andup to four non-aromatic carbon-carbon double bonds may be present. Thus,“C₂-C₆ alkenyl” means an alkenyl radical having from 2 to 6 carbonatoms. Alkenyl groups include ethenyl, propenyl, butenyl,2-methylbutenyl and cyclohexenyl. The straight, branched or cyclicportion of the alkenyl group may contain double bonds and may besubstituted if a substituted alkenyl group is indicated.

The term “alkynyl” refers to a hydrocarbon radical straight, branched orcyclic, containing from 2 to 10 carbon atoms and at least one carbon tocarbon triple bond. Up to three carbon-carbon triple bonds may bepresent. Thus, “C₂-C₆ alkynyl” means an alkynyl radical having from 2 to6 carbon atoms. Alkynyl groups include ethynyl, propynyl, butynyl,3-methylbutynyl and so on. The straight, branched or cyclic portion ofthe alkynyl group may contain triple bonds and may be substituted if asubstituted alkynyl group is indicated.

In certain instances, substituents may be defined with a range ofcarbons that includes zero, such as (C₀-C₆)alkylene-aryl. If aryl istaken to be phenyl, this definition would include phenyl itself as wellas —CH₂Ph, —CH₂CH₂Ph, —CH(CH₃)CH₂CH(CH₃)Ph, and so on.

As used herein, “aryl” is intended to mean any stable monocyclic orbicyclic carbon ring of up to 7 atoms in each ring, wherein at least onering is aromatic. Examples of such aryl elements include phenyl,naphthyl, tetrahydro-naphthyl, indanyl and biphenyl. In cases where thearyl substituent is bicyclic and one ring is non-aromatic, it isunderstood that attachment is via the aromatic ring.

The term heteroaryl, as used herein, represents a stable monocyclic orbicyclic ring of up to 7 atoms in each ring, wherein at least one ringis aromatic and contains from 1 to 4 heteroatoms selected from the groupconsisting of O, N and S. Heteroaryl groups within the scope of thisdefinition include but are not limited to: acridinyl, carbazolyl,cinnolinyl, quinoxalinyl, pyrrazolyl, indolyl, benzotriazolyl, furanyl,thienyl, benzothienyl, benzofuranyl, quinolinyl, isoquinolinyl,oxazolyl, isoxazolyl, indolyl, pyrazinyl, pyridazinyl, pyridinyl,pyrimidinyl, pyrrolyl, tetrahydroquinoline. As with the definition ofheterocycle below, “heteroaryl” is also understood to include theN-oxide derivative of any nitrogen-containing heteroaryl. In cases wherethe heteroaryl substituent is bicyclic and one ring is non-aromatic orcontains no heteroatoms, it is understood that attachment is via thearomatic ring or via the heteroatom containing ring, respectively. Suchheteraoaryl moieties for substituent Q include but are not limited to:2-benzimidazolyl, 2-quinolinyl, 3-quinolinyl, 4-quinolinyl,1-isoquinolinyl, 3-isoquinolinyl and 4-isoquinolinyl.

The term “heterocycle” or “heterocyclyl” as used herein is intended tomean a 3- to 10-membered aromatic or nonaromatic heterocycle containingfrom 1 to 4 heteroatoms selected from the group consisting of O, N andS, and includes bicyclic groups. “Heterocyclyl” therefore includes theabove mentioned heteroaryls, as well as dihydro and tetrathydro analogsthereof. Further examples of “heterocyclyl” include, but are not limitedto the following: benzoimidazolyl, benzoimidazolonyl, benzofuranyl,benzofurazanyl, benzopyrazolyl, benzotriazolyl, benzothiophenyl,benzoxazolyl, carbazolyl, carbolinyl, cinnolinyl, furanyl, imidazolyl,indolinyl, indolyl, indolazinyl, indazolyl, isobenzofuranyl, isoindolyl,isoquinolyl, isothiazolyl, isoxazolyl, naphthpyridinyl, oxadiazolyl,oxazolyl, oxazoline, isoxazoline, oxetanyl, pyranyl, pyrazinyl,pyrazolyl, pyridazinyl, pyridopyridinyl, pyridazinyl,pyrazolopyrimidinyl, pyridyl, pyrimidyl, pyrrolyl, quinazolinyl,quinolyl, quinoxalinyl, tetrahydropyranyl, tetrazolyl, tetrazolopyridyl,thiadiazolyl, thiazolyl, thienyl, triazolyl, azetidinyl, 1,4-dioxanyl,hexahydroazepinyl, piperazinyl, piperidinyl, pyridin-2-onyl,pyrrolidinyl, morpholinyl, thiomorpholinyl, dihydrobenzoimidazolyl,dihydrobenzofuranyl, dihydrobenzothiophenyl, dihydrobenzoxazolyl,dihydrofuranyl, dihydroimidazolyl, dihydroindolyl, dihydroisooxazolyl,dihydroisothiazolyl, dihydrooxadiazolyl, dihydrooxazolyl,dihydropyrazinyl, dihydropyrazolyl, dihydropyridinyl,dihydropyrimidinyl, dihydropyrrolyl, dihydroquinolinyl,dihydrotetrazolyl, dihydrothiadiazolyl, dihydrothiazolyl,dihydrothienyl, dihydrotriazolyl, dihydroazetidinyl,methylenedioxybenzoyl, tetrahydrofuranyl, and tetrahydrothienyl, andN-oxides thereof. Attachment of a heterocyclyl substituent can occur viaa carbon atom or via a heteroatom.

As appreciated by those of skill in the art, “halo” or “halogen” as usedherein is intended to include chloro, fluoro, bromo and iodo.

As used herein, unless otherwise specifically defined, substitutedalkyl, substituted cycloalkyl, substituted aryl, substituted heteroaryl,substituted arylsulfonyl, substituted heteroaryl-sulfonyl andsubstituted heterocycle include moieties containing from 1 to 4substituents (and in another embodiment 1 to 3 substituents) in additionto the point of attachment to the rest of the compound. Suchsubstituents are selected from the group which includes but is notlimited to F, Cl, Br, CF₃, NH₂, N(C₁-C₆ alkyl)₂, NO₂, CN, (C₁-C₆alkyl)O—, (aryl)O—, —OH, O_(a)-P═O(OH)₂, (C₁-C₆ alkyl)S(O)_(m)—, (C₁-C₆alkyl)C(O)NH—, H₂N—C(NH)—, (C₁-C₆ alkyl)C(O)—, (C₁-C₆ alkyl)OC(O)—,(C₁-C₆ alkyl)OC(O)NH—, phenyl, pyridyl, imidazolyl, oxazolyl,isoxazolyl, tetrazolyl, thiazolyl, thienyl, furyl, isothiazolyl andC₁-C₂₀ alkyl. For example, a (C₁-C₆)alkyl may be substituted with one,two, three or four (in another embodiment one, two or three)substituents selected from OH, oxo, halogen, alkoxy, dialkylamino, orheterocyclyl, such as morpholinyl, piperidinyl, and so on. In this case,if one substituent is oxo and the other is OH, the following areincluded in the definition: —(C═O)CH₂CH(OH)CH₃, —(C═O)OH,—CH₂(OH)CH₂CH(O), and so on.

In another embodiment, n is 0 or 1.

In a further embodiment, n is 1.

In another embodiment, R^(a) is H, (C₁-C₆)alkyl or phenyl.

In another embodiment, R^(b) is independently H, (C₁-C₆)alkyl,(C═O)O(C₁-C₆)alkyl, (C═O)(C₁-C₆)alkyl or S(O)₂R^(a). In an embodiment, Qis selected from 2-azepinone, benzimidazolyl, benzimidazolonyl,2-diazapinone, imidazolyl, 2-imidazolidinone, indolyl, isoquinolinyl,morpholinyl, piperidyl, piperazinyl, pyridyl, pyrrolidinyl,2-piperidinone, 2-pyrimidinone, 2-pyrollidinone, quinolinyl, tetrazolyl,tetrahydrofuryl, tetrahydroisoquinolinyl, thienyl, pyrazolopyrimidinyl,pyrazolyl, thiazolyl, oxadiazolyl and triazolyl, optionally substitutedwith 1-3 R^(z).

In a further embodiment, Q is selected from:

which are optionally substituted with one to three substituents selectedfrom R^(z).

In still a further embodiment, Q is selected from

which are optionally substituted with one substituent selected fromR^(z).

In a further embodiment, Q is selected from

which are optionally substituted with one substituent selected fromR^(z).

In yet a further embodiment, Q is selected from:

which are optionally substituted with one substituent selected fromR^(z).

In another embodiment, R^(z) is selected from: (C₁-C₆)alkyl, phenyl,heterocyclyl, N(R^(b))₂, (C═O)_(a)O₆C₁-C₆allyl, (C═O)NR²R³, halogen, OH,O_(a)-P═O(OH)₂, and oxo.

In a further embodiment, R^(z) is selected from: (C₁-C₆)alkyl, phenyl,heterocyclyl, N(R^(b))₂, (C═O)_(a)O₆C₁-C₆alkyl, (C═O)NR²R³, halogen, OH,O_(a)-P═O(OH)₂, and oxo, wherein said heterocyclyl is selected from:

In still a further embodiment, when R^(z) is heterocyclyl, saidheterocyclyl is selected from:

In yet a further embodiment, R^(z) is selected from: NH₂ andheterocyclyl, said heterocyclyl is selected from

In yet a further embodiment, R^(z) is:

In another embodiment, R¹ is selected from: OH; heterocyclyl; andNR^(a)R^(b); wherein R^(a) and R^(b) are independently H or (C₁-C₆)alkyland wherein said heterocyclyl is optionally substituted with oxo and(C═O)_(r)O_(s)(C₁-C₆)alkyl.

In another embodiment, when R¹ is heterocyclyl, said heterocyclyl isselected from:

In a further embodiment, when R¹ is heterocyclyl, said heterocyclyl isselected from:

In another embodiment, R² and R³ are independently selected from H,(C₁-C₆)alkyl and aryl, optionally substituted with one to twosubstituents selected from R^(z), or R² and R³ together with thenitrogen to which they are attached form a monocyclic or bicyclicheterocycle, optionally substituted with one to two substituentsselected from R^(z).

In a further embodiment, R² and R³ are independently selected from H or(C₁-C₆)alkyl, optionally substituted with one to two substituentsselected from R^(z), or R² and R³ together with the nitrogen to whichthey are attached form a monocyclic or bicyclic heterocycle, optionallysubstituted with one to two substituents selected from R^(z).

In a fourth embodiment of this invention, the inhibitors of Akt activityare illustrated by the Formula A-3:

wherein:

X is N and Y is CH or X is CH and Y is N;

Q is: R¹;

a is 0 or 1; b is 0 or 1; m is 0, 1, or 2; n is 0, 1, 2, 3 or 4; p is 0,1, 2, 3, 4 or 5;

R¹ is independently selected from: H, (C═O)_(a)O_(b)C₁-C₁₀ alkyl,(C═O)_(a)O_(b) aryl, (C═O)_(a)O_(b)C₂-C₁₀ alkenyl, (C═O)_(a)O_(b)C₂-C₁₀alkynyl, CO₂H, halo, OH, oxo, O_(b)C₁-C₆ perfluoroalkyl, (C═O)_(a)NR⁷R⁸,CN, (C═O)_(a)O_(b)C₃-C₈ cycloalkyl, S(O)_(m)NR⁷R⁸,S(O)_(m)—(C₁-C₁₀)alkyl and (C═O)_(a)O_(b)heterocyclyl, said alkyl, aryl,alkenyl, alkynyl, cycloalkyl, and heterocyclyl is optionally substitutedwith one or more substituents selected from R⁶;

R²′ is independently selected from: H, (C═O)_(a)O_(b)C₁-C₁₀ alkyl,(C═O)_(a)O_(b) aryl, (C═O)_(a)O_(b)C₂-C₁₀ alkenyl, (C═O)_(a)O_(b)C₂-C₁₀alkynyl, CO₂H, halo, OH, oxo, O_(b)C₁-C₆ perfluoroalkyl, (C═O)_(a)NR⁷R⁸,CN, (C═O)_(a)O_(b)C₃-C₈ cycloalkyl, S(O)_(m)NR⁷R⁸,S(O)_(m)—(C₁-C₁₀)alkyl and (C═O)_(a)O_(b)heterocyclyl, said alkyl, aryl,alkenyl, alkynyl, cycloalkyl, and heterocyclyl is optionally substitutedwith one or more substituents selected from R⁶;

R⁶ is independently: (C═O)_(a)O_(b)C₁-C₁₀ alkyl, (C═O)_(a)O_(b)aryl,C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, (C═O)_(a)O_(b) heterocyclyl, CO₂H, halo,CN, OH, O_(b)C₁-C₆ perfluoroalkyl, O_(a)(C═O)_(b)NR⁷R⁸, oxo, CHO,(N═O)R⁷R⁸, S(O)_(m)NR⁷R⁸, S(O)_(m)—(C₁-C₁₀) alkyl or (C═O)_(a)O_(b)C₃-C₈cycloalkyl, said alkyl, aryl, alkenyl, alkynyl, heterocyclyl, andcycloalkyl optionally substituted with one or more substituents selectedfrom R^(6a);

R^(6a) is independently selected from: (C═O)_(a)O_(b)(C₁-C₁₀)alkyl,O_(a)(C₁-C₃)perfluoroalkyl, (C₀-C₆)alkylene-S(O)_(m)R^(a), oxo, OH,halo, CN, (C₂-C₁₀)alkenyl, (C₂-C₁₀)alkynyl, (C₃-C₆)cycloalkyl,(C₀-C₆)alkylene-aryl, (C₀-C₆)alkylene-heterocyclyl,(C₀-C₆)alkylene-N(R^(b))₂, C(O)R^(a), (C₀-C₆)alkylene-CO₂R^(a), C(O)H,and (C₀-C₆)alkylene-CO₂H, said alkyl, alkenyl, alkynyl, cycloalkyl,aryl, and heterocyclyl is optionally substituted with up to threesubstituents selected from R^(b), OH, (C₁-C₆)alkoxy, halogen, CO₂H, CN,O(C═O)C₁-C₆ alkyl, oxo, and N(R^(b))₂;

R⁷ and R⁸ are independently selected from: H, (C═O)O_(b)C₁-C₁₀ alkyl,(C═O)O_(b)C₃-C₈ cycloalkyl, (C═O)O_(b)aryl, (C═O)O_(b)heterocyclyl,C₁-C₁₀ alkyl, aryl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, heterocyclyl, C₃-C₉cycloalkyl, SO₂R^(a), and (C═O)NR^(b) ₂, said alkyl, cycloalkyl, aryl,heterocylyl, alkenyl, and alkynyl is optionally substituted with one ormore substituents selected from R^(6a), or R⁷ and R⁸ can be takentogether with the nitrogen to which they are attached to form amonocyclic or bicyclic heterocycle with 5-7 members in each ring andoptionally containing, in addition to the nitrogen, one or twoadditional heteroatoms selected from N, O and S, said monocylcic orbicyclic heterocycle optionally substituted with one or moresubstituents selected from R^(6a);

R^(a)′ is independently: (C₁-C₆)alkyl, (C₃-C₆)cycloalkyl, aryl, orheterocyclyl; and

R^(b)′ is independently: H, (C₁-C₆)alkyl, aryl, heterocyclyl,(C₃-C₆)cycloalkyl, (C═O)OC₁-C₆ alkyl, (C═O)C₁-C₆ alkyl or S(O)₂R^(a);

or a pharmaceutically acceptable salt or a stereoisomer thereof.

In a fifth embodiment of this invention, the inhibitors of Akt activityare illustrated by the Formula A-3:

wherein:

Q is: (C₃-C₈)cycloalkyl, aryl or heterocyclyl, which is optionallysubstituted with 1-3 R⁶;

and all other substituents and variables are as defined in the fourthembodiment;

or a pharmaceutically acceptable salt or a stereoisomer thereof.

In a sixth embodiment of this invention, the inhibitors of Akt activityare illustrated by the Formula B-3:

wherein:

Q is selected from:

which is optionally substituted with 1-3 R⁶;

n is 0, 1 or 2;

and all other substituents and variables are as defined in the fifthembodiment;

or a pharmaceutically acceptable salt or a stereoisomer thereof.

In a seventh embodiment of this invention, the inhibitors of Aktactivity are illustrated by the Formula B-3: wherein:

Q is selected from

which are optionally substituted with 1-3 R⁶;

and all other substituents and variables are as defined in the sixthembodiment;

or a pharmaceutically acceptable salt or a stereoisomer thereof.

In an embodiment, when R⁶ is attached to Q; R⁶ is independently selectedfrom: (C₁-C₆)alkyl, phenyl, heterocyclyl, N(R^(b))₂,(C═O)_(a)O_(b)C₁-C₆alkyl, (C═O)NR²R³, halogen, OH, O_(a)-P═O(OH)₂, andoxo.

In an embodiment, when R⁶ is attached to Q; R⁶ is independently selectedfrom:

Included in the instant invention is the free form of compounds ofFormula A, as well as the pharmaceutically acceptable salts andstereoisomers thereof. Some of the isolated specific compoundsexemplified herein are the protonated salts of amine compounds. The term“free form” refers to the amine compounds in non-salt form. Theencompassed pharmaceutically acceptable salts not only include theisolated salts exemplified for the specific compounds described herein,but also all the typical pharmaceutically acceptable salts of the freeform of compounds of Formula A. The free form of the specific saltcompounds described may be isolated using techniques known in the art.For example, the free form may be regenerated by treating the salt witha suitable dilute aqueous base solution such as dilute aqueous NaOH,potassium carbonate, ammonia and sodium bicarbonate. The free forms maydiffer from their respective salt forms somewhat in certain physicalproperties, such as solubility in polar solvents, but the acid and basesalts are otherwise pharmaceutically equivalent to their respective freeforms for purposes of the invention.

The pharmaceutically acceptable salts of the instant compounds can besynthesized from the compounds of this invention which contain a basicor acidic moiety by conventional chemical methods. Generally, the saltsof the basic compounds are prepared either by ion exchangechromatography or by reacting the free base with stoichiometric amountsor with an excess of the desired salt-forming inorganic or organic acidin a suitable solvent or various combinations of solvents. Similarly,the salts of the acidic compounds are formed by reactions with theappropriate inorganic or organic base.

Thus, pharmaceutically acceptable salts of the compounds of thisinvention include the conventional non-toxic salts of the compounds ofthis invention as formed by reacting a basic instant compound with aninorganic or organic acid. For example, conventional non-toxic saltsinclude those derived from inorganic acids such as hydrochloric,hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like, aswell as salts prepared from organic acids such as acetic, propionic,succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic,pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic,salicylic, sulfanilic, 2-acetoxy-benzoic, fumaric, toluenesulfonic,methanesulfonic, ethane disulfonic, oxalic, isethionic, trifluoroacetic(TFA) and the like.

When the compound of the present invention is acidic, suitable“pharmaceutically acceptable salts” refers to salts prepared formpharmaceutically acceptable non-toxic bases including inorganic basesand organic bases. Salts derived from inorganic bases include aluminum,ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganicsalts, manganous, potassium, sodium, zinc and the like. Particularlypreferred are the ammonium, calcium, magnesium, potassium and sodiumsalts. Salts derived from pharmaceutically acceptable organic non-toxicbases include salts of primary, secondary and tertiary amines,substituted amines including naturally occurring substituted amines,cyclic amines and basic ion exchange resins, such as arginine, betainecaffeine, choline, N,N¹-dibenzylethylenediamine, diethylamine,2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine,ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine,glucosamine, histidine, hydrabamine, isopropylamine, lysine,methylglucamine, morpholine, piperazine, piperidine, polyamine resins,procaine, purines, theobromine, triethylamine, trimethylaminetripropylamine, tromethamine and the like.

The preparation of the pharmaceutically acceptable salts described aboveand other typical pharmaceutically acceptable salts is more fullydescribed by Berg et al., “Pharmaceutical Salts,” J. Pharm. Sci.,1977:66:1-19.

It will also be noted that the compounds of the present invention arepotentially internal salts or zwitterions, since under physiologicalconditions a deprotonated acidic moiety in the compound, such as acarboxyl group, may be anionic, and this electronic charge might then bebalanced off internally against the cationic charge of a protonated oralkylated basic moiety, such as a quaternary nitrogen atom.

Utility

The compounds of the instant invention are inhibitors of the activity ofAkt and are thus useful in the treatment of cancer, in particularcancers associated with irregularities in the activity of Akt anddownstream cellular targets of Akt. Such cancers include, but are notlimited to, ovarian, pancreatic, breast and prostate cancer, as well ascancers (including glioblastoma) where the tumor suppressor PTEN ismutated (Cheng et al., Proc. Natl. Acad. Sci. (1992) 89:9267-9271; Chenget al., Proc. Natl. Acad. Sci. (1996) 93:3636-3641; Bellacosa et al.,Int. J. Cancer (1995) 64:280-285; Nakatani et al., J. Biol. Chem. (1999)274:21528-21532; Graff, Expert. Opin. Ther. Targets (2002) 6(1):103-113;and Yamada and Araki, J. Cell Science. (2001) 114:2375-2382; Mischel andCloughesy, Brain Pathol. (2003) 13(1):52-61).

The compounds, compositions and methods provided herein are particularlydeemed useful for the treatment of cancer including solid tumors such asskin, breast, brain, cervical carcinomas, testicular carcinomas, etc.More particularly, cancers that may be treated by the compounds,compositions and methods of the invention include, but are not limitedto: Cardiac: sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma,liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma and teratoma; Lung:bronchogenic carcinoma (squamous cell, undifferentiated small cell,undifferentiated large cell, adenocarcinoma), alveolar (bronchiolar)carcinoma, bronchial adenoma, sarcoma, lymphoma, chondromatoushamartoma, mesothelioma; Gastrointestinal: esophagus (squamous cellcarcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach(carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma,insulinoma, glucagonoma, gastrinoma, carcinoid tumors, vipoma), smallbowel (adenocarcinoma, lymphoma, carcinoid tumors, Karposi's sarcoma,leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large bowel(adenocarcinoma, tubular adenoma, villous adenoma, hamartoma,leiomyoma); Genitourinary tract: kidney (adenocarcinoma, Wilm's tumor[nephroblastoma], lymphoma, leukemia), bladder and urethra (squamouscell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate(adenocarcinoma, sarcoma), testis (seminoma, teratoma, embryonalcarcinoma, teratocarcinoma, choriocarcinoma, sarcoma, interstitial cellcarcinoma, fibroma, fibroadenoma, adenomatoid tumors, lipoma); Liver:hepatoma (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma,angiosarcoma, hepatocellular adenoma, hemangioma; Bone: osteogenicsarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma,chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulum cellsarcoma), multiple myeloma, malignant giant cell tumor chordoma,osteochronfroma (osteocartilaginous exostoses), benign chondroma,chondroblastoma, chondromyxofibroma, osteoid osteoma and giant celltumors; Nervous system: skull (osteoma, hemangioma, granuloma, xanthoma,osteitis deformans), meninges (meningioma, meningiosarcoma,gliomatosis), brain (astrocytoma, medulloblastoma, glioma, ependymoma,germinoma [pinealoma], glioblastoma multiform, oligodendroglioma,schwannoma, retinoblastoma, congenital tumors), spinal cordneurofibroma, meningioma, glioma, sarcoma); Gynecological: uterus(endometrial carcinoma), cervix (cervical carcinoma, pre-tumor cervicaldysplasia), ovaries (ovarian carcinoma [serous cystadenocarcinoma,mucinous cystadenocarcinoma, unclassified carcinoma], granulosa-thecalcell tumors, Sertoli-Leydig cell tumors, dysgerminoma, malignantteratoma), vulva (squamous cell carcinoma, intraepithelial carcinoma,adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma,squamous cell carcinoma, botryoid sarcoma (embryonal rhabdomyosarcoma),fallopian tubes (carcinoma); Hematologic: blood (myeloid leukemia [acuteand chronic], acute lymphoblastic leukemia, chronic lymphocyticleukemia, myeloproliferative diseases, multiple myeloma, myelodysplasticsyndrome), Hodgkin's disease, non-Hodgkin's lymphoma [malignantlymphoma]; Skin: malignant melanoma, basal cell carcinoma, squamous cellcarcinoma, Karposi's sarcoma, moles dysplastic nevi, lipoma, angioma,dermatofibroma, keloids, psoriasis; and Adrenal glands: neuroblastoma.Thus, the term “cancerous cell” as provided herein, includes a cellafflicted by any one of the above-identified conditions.

Akt signaling regulates multiple critical steps in angiogenesis.Shiojima and Walsh, Circ. Res. (2002) 90:1243-1250. The utility ofangiogenesis inhibitors in the treatment of cancer is known in theliterature, see J. Rak et al. Cancer Research, 55:4575-4580, 1995 andDredge et al., Expert Opin. Biol. Ther. (2002) 2(8):953-966, forexample. The role of angiogenesis in cancer has been shown in numeroustypes of cancer and tissues: breast carcinoma (G. Gasparini and A. L.Harris, J. Clin. Oncol., 1995, 13:765-782; M. Toi et al., Japan. J.Cancer Res., 1994, 85:1045-1049); bladder carcinomas (A. J. Dickinson etal., Br. J. Urol., 1994, 74:762-766); colon carcinomas (L. M. Ellis etal., Surgery, 1996, 120(5):871-878); and oral cavity tumors (J. K.Williams et al., Am. J. Surg., 1994, 168:373-380). Other cancersinclude, advanced tumors, hairy cell leukemia, melanoma, advanced headand neck, metastatic renal cell, non-Hodgkin's lymphoma, metastaticbreast, breast adenocarcinoma, advanced melanoma, pancreatic, gastric,glioblastoma, lung, ovarian, non-small cell lung, prostate, small celllung, renal cell carcinoma, various solid tumors, multiple myeloma,metastatic prostate, malignant glioma, renal cancer, lymphoma,refractory metastatic disease, refractory multiple myeloma, cervicalcancer, Kaposi's sarcoma, recurrent anaplastic glioma, and metastaticcolon cancer (Dredge et al., Expert Opin. Biol. Ther. (2002)2(8):953-966). Thus, the Akt inhibitors disclosed in the instantapplication, are also useful in the treatment of these angiogenesisrelated cancers.

Tumors which have undergone neovascularization show an increasedpotential for metastasis. In fact, angiogenesis is essential for tumorgrowth and metastasis. (S. P. Cunningham, et al., Can. Research, 61:3206-3211 (2001)). The Akt inhibitors disclosed in the presentapplication are therefore also useful to prevent or decrease tumor cellmetastasis.

Further included within the scope of the invention is a method oftreating or preventing a disease in which angiogenesis is implicated,which is comprised of administering to a mammal in need of suchtreatment a therapeutically effective amount of a compound of thepresent invention. Ocular neovascular diseases are an example ofconditions where much of the resulting tissue damage can be attributedto aberrant infiltration of blood vessels in the eye (see WO 00/30651,published 2 Jun. 2000). The undesirable infiltration can be triggered byischemic retinopathy, such as that resulting from diabetic retinopathy,retinopathy of prematurity, retinal vein occlusions, etc., or bydegenerative diseases, such as the choroidal neovascularization observedin age-related macular degeneration. Inhibiting the growth of bloodvessels by administration of the present compounds would thereforeprevent the infiltration of blood vessels and prevent or treat diseaseswhere angiogenesis is implicated, such as ocular diseases like retinalvascularization, diabetic retinopathy, age-related macular degeneration,and the like.

Further included within the scope of the invention is a method oftreating or preventing a non-malignant disease in which angiogenesis isimplicated, including but not limited to: ocular diseases (such as,retinal vascularization, diabetic retinopathy and age-related maculardegeneration), atherosclerosis, arthritis, psoriasis, obesity andAlzheimer's disease (Dredge et al., Expert Opin. Biol. Ther. (2002)2(8):953-966). In another embodiment, a method of treating or preventinga disease in which angiogenesis is implicated includes: ocular diseases(such as, retinal vascularization, diabetic retinopathy and age-relatedmacular degeneration), atherosclerosis, arthritis and psoriasis.

Further included within the scope of the invention is a method oftreating hyperproliferative disorders such as restenosis, inflammation,autoimmune diseases and allergy/asthma.

Further included within the scope of the invention is a method oftreating hyperinsulinism.

The compounds of the invention are also useful in preparing a medicamentthat is useful in treating the diseases described above, in particularcancer.

In an embodiment of the invention, the instant compound is a selectiveinhibitor whose inhibitory efficacy is dependent on the PH domain. Inthis embodiment, the compound exhibits a decrease in in vitro inhibitoryactivity or no in vitro inhibitory activity against truncated Aktproteins lacking the PH domain.

In a further embodiment, the instant compound is selected from the groupof a selective inhibitor of Akt1, a selective inhibitor of Akt2 and aselective inhibitor of both Akt1 and Akt2.

In another embodiment, the instant compound is selected from the groupof a selective inhibitor of Akt1, a selective inhibitor of Akt2, aselective inhibitor of Akt3 and a selective inhibitor of two of thethree Akt isoforms.

In another embodiment, the instant compound is a selective inhibitor ofall three Akt isoforms, but is not an inhibitor of one, two or all ofsuch Akt isoforms that have been modified to delete the PH domain, thehinge region or both the PH domain and the hinge region.

The present invention is further directed to a method of inhibiting Aktactivity which comprises administering to a mammal in need thereof apharmaceutically effective amount of the instant compound.

The compounds of this invention may be administered to mammals,including humans, either alone or, in combination with pharmaceuticallyacceptable carriers, excipients or diluents, in a pharmaceuticalcomposition, according to standard pharmaceutical practice. Thecompounds can be administered orally or parenterally, including theintravenous, intramuscular, intraperitoneal, subcutaneous, rectal andtopical routes of administration.

The pharmaceutical compositions containing the active ingredient may bein a form suitable for oral use, for example, as tablets, troches,lozenges, aqueous or oily suspensions, dispersible powders or granules,emulsions, hard or soft capsules, or syrups or elixirs. Compositionsintended for oral use may be prepared according to any method known tothe art for the manufacture of pharmaceutical compositions and suchcompositions may contain one or more agents selected from the groupconsisting of sweetening agents, flavoring agents, coloring agents andpreserving agents in order to provide pharmaceutically elegant andpalatable preparations. Tablets contain the active ingredient inadmixture with non-toxic pharmaceutically acceptable excipients whichare suitable for the manufacture of tablets. These excipients may be forexample, inert diluents, such as calcium carbonate, sodium carbonate,lactose, calcium phosphate or sodium phosphate; granulating anddisintegrating agents, for example, microcrystalline cellulose, sodiumcrosscarmellose, corn starch, or alginic acid; binding agents, forexample starch, gelatin, polyvinyl-pyrrolidone or acacia, andlubricating agents, for example, magnesium stearate, stearic acid ortalc. The tablets may be uncoated or they may be coated by knowntechniques to mask the unpleasant taste of the drug or delaydisintegration and absorption in the gastrointestinal tract and therebyprovide a sustained action over a longer period. For example, a watersoluble taste masking material such as hydroxypropylmethyl-cellulose orhydroxypropylcellulose, or a time delay material such as ethylcellulose, cellulose acetate buryrate may be employed.

Formulations for oral use may also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules wherein the active ingredient is mixed with watersoluble carrier such as polyethyleneglycol or an oil medium, for examplepeanut oil, liquid paraffin, or olive oil.

Aqueous suspensions contain the active material in admixture withexcipients suitable for the manufacture of aqueous suspensions. Suchexcipients are suspending agents, for example sodiumcarboxymethylcellulose, methylcellulose, hydroxypropylmethyl-cellulose,sodium alginate, polyvinyl-pyrrolidone, gum tragacanth and gum acacia;dispersing or wetting agents may be a naturally-occurring phosphatide,for example lecithin, or condensation products of an alkylene oxide withfatty acids, for example polyoxyethylene stearate, or condensationproducts of ethylene oxide with long chain aliphatic alcohols, forexample heptadecaethylene-oxycetanol, or condensation products ofethylene oxide with partial esters derived from fatty acids and ahexitol such as polyoxyethylene sorbitol monooleate, or condensationproducts of ethylene oxide with partial esters derived from fatty acidsand hexitol anhydrides, for example polyethylene sorbitan monooleate.The aqueous suspensions may also contain one or more preservatives, forexample ethyl, or n-propyl p-hydroxybenzoate, one or more coloringagents, one or more flavoring agents, and one or more sweetening agents,such as sucrose, saccharin or aspartame.

Oily suspensions may be formulated by suspending the active ingredientin a vegetable oil, for example arachis oil, olive oil, sesame oil orcoconut oil, or in mineral oil such as liquid paraffin. The oilysuspensions may contain a thickening agent, for example beeswax, hardparaffin or cetyl alcohol. Sweetening agents such as those set forthabove, and flavoring agents may be added to provide a palatable oralpreparation. These compositions may be preserved by the addition of ananti-oxidant such as butylated hydroxyanisol or alpha-tocopherol.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water provide the active ingredient inadmixture with a dispersing or wetting agent, suspending agent and oneor more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.Additional excipients, for example sweetening, flavoring and coloringagents, may also be present. These compositions may be preserved by theaddition of an anti-oxidant such as ascorbic acid.

The pharmaceutical compositions of the invention may also be in the formof an oil-in-water emulsion. The oily phase may be a vegetable oil, forexample olive oil or arachis oil, or a mineral oil, for example liquidparaffin or mixtures of these. Suitable emulsifying agents may benaturally-occurring phosphatides, for example soy bean lecithin, andesters or partial esters derived from fatty acids and hexitolanhydrides, for example sorbitan monooleate, and condensation productsof the said partial esters with ethylene oxide, for examplepolyoxyethylene sorbitan monooleate. The emulsions may also containsweetening, flavouring agents, preservatives and antioxidants.

Syrups and elixirs may be formulated with sweetening agents, for exampleglycerol, propylene glycol, sorbitol or sucrose. Such formulations mayalso contain a demulcent, a preservative, flavoring and coloring agentsand antioxidant.

The pharmaceutical compositions may be in the form of sterile injectableaqueous solutions. Among the acceptable vehicles and solvents that maybe employed are water, Ringer's solution and isotonic sodium chloridesolution.

The sterile injectable preparation may also be a sterile injectableoil-in-water microemulsion where the active ingredient is dissolved inthe oily phase. For example, the active ingredient may be firstdissolved in a mixture of soybean oil and lecithin. The oil solutionthen introduced into a water and glycerol mixture and processed to forma microemulation.

The injectable solutions or microemulsions may be introduced into apatient's blood-stream by local bolus injection. Alternatively, it maybe advantageous to administer the solution or microemulsion in such away as to maintain a constant circulating concentration of the instantcompound. In order to maintain such a constant concentration, acontinuous intravenous delivery device may be utilized. An example ofsuch a device is the Deltec CADD-PLUS™ model 5400 intravenous pump.

The pharmaceutical compositions may be in the form of a sterileinjectable aqueous or oleagenous suspension for intramuscular andsubcutaneous administration. This suspension may be formulated accordingto the known art using those suitable dispersing or wetting agents andsuspending agents which have been mentioned above. The sterileinjectable preparation may also be a sterile injectable solution orsuspension in a non-toxic parenterally-acceptable diluent or solvent,for example as a solution in 1,3-butane diol. In addition, sterile,fixed oils are conventionally employed as a solvent or suspendingmedium. For this purpose any bland fixed oil may be employed includingsynthetic mono- or diglycerides. In addition, fatty acids such as oleicacid find use in the preparation of injectables.

Compounds of Formula A may also be administered in the form ofsuppositories for rectal administration of the drug. These compositionscan be prepared by mixing the drug with a suitable non-irritatingexcipient which is solid at ordinary temperatures but liquid at therectal temperature and will therefore melt in the rectum to release thedrug. Such materials include cocoa butter, glycerinated gelatin,hydrogenated vegetable oils, mixtures of polyethylene glycols of variousmolecular weights and fatty acid esters of polyethylene glycol.

For topical use, creams, ointments, jellies, solutions or suspensions,etc., containing the compound of Formula A are employed. (For purposesof this application, topical application shall include mouth washes andgargles.)

The compounds for the present invention can be administered inintranasal form via topical use of suitable intranasal vehicles anddelivery devices, or via transdermal routes, using those forms oftransdermal skin patches well known to those of ordinary skill in theart. To be administered in the form of a transdermal delivery system,the dosage administration will, of course, be continuous rather thanintermittent throughout the dosage regimen. Compounds of the presentinvention may also be delivered as a suppository employing bases such ascocoa butter, glycerinated gelatin, hydrogenated vegetable oils,mixtures of polyethylene glycols of various molecular weights and fattyacid esters of polyethylene glycol.

When a composition according to this invention is administered into ahuman subject, the daily dosage will normally be determined by theprescribing physician with the dosage generally varying according to theage, weight, and response of the individual patient, as well as theseverity of the patient's symptoms.

In an embodiment, a suitable amount of an inhibitor of Akt isadministered to a mammal undergoing treatment for cancer. Administrationoccurs in an amount of inhibitor of between about 0.1 mg/kg of bodyweight to about 60 mg/kg of body weight per day, or between 0.5 mg/kg ofbody weight to about 40 mg/kg of body weight per day. Anothertherapeutic dosage that comprises the instant composition includes fromabout 0.01 mg to about 1000 mg of inhibitor of Akt. In anotherembodiment, the dosage comprises from about 1 mg to about 1000 mg ofinhibitor of Akt.

The instant compounds are also useful in combination with knowntherapeutic agents and anti-cancer agents. For example, instantcompounds are useful in combination with known anti-cancer agents.Combinations of the presently disclosed compounds with other anti-canceror chemotherapeutic agents are within the scope of the invention.Examples of such agents can be found in Cancer Principles and Practiceof Oncology by V. T. Devita and S. Hellman (editors), 6^(th) edition(Feb. 15, 2001), Lippincott Williams & Wilkins Publishers. A person ofordinary skill in the art would be able to discern which combinations ofagents would be useful based on the particular characteristics of thedrugs and the cancer involved. Such anti-cancer agents include thefollowing: estrogen receptor modulators, androgen receptor modulators,retinoid receptor modulators, cytotoxic/cytostatic agents,antiproliferative agents, prenyl-protein transferase inhibitors, HMG-CoAreductase inhibitors and other angiogenesis inhibitors, inhibitors ofcell proliferation and survival signaling, and agents that interferewith cell cycle checkpoints. The instant compounds are particularlyuseful when co-administered with radiation therapy.

In an embodiment, the instant compounds are also useful in combinationwith known anti-cancer agents including the following: estrogen receptormodulators, androgen receptor modulators, retinoid receptor modulators,cytotoxic agents, antiproliferative agents, prenyl-protein transferaseinhibitors, HMG-CoA reductase inhibitors, HIV protease inhibitors,reverse transcriptase inhibitors, and other angiogenesis inhibitors.

“Estrogen receptor modulators” refers to compounds that interfere withor inhibit the binding of estrogen to the receptor, regardless ofmechanism. Examples of estrogen receptor modulators include, but are notlimited to, tamoxifen, raloxifene, idoxifene, LY353381, LY117081,toremifene, fulvestrant,4-[7-(2,2-dimethyl-1-oxopropoxy-4-methyl-2-[4-[2-(1-piperidinyl)ethoxy]phenyl]-2H-1-benzopyran-3-yl]-phenyl-2,2-dimethylpropanoate,4,4′-dihydroxybenzophenone-2,4-dinitrophenyl-hydrazone, and SH646.

“Androgen receptor modulators” refers to compounds which interfere orinhibit the binding of androgens to the receptor, regardless ofmechanism. Examples of androgen receptor modulators include finasterideand other 5α-reductase inhibitors, nilutamide, flutamide, bicalutamide,liarozole, and abiraterone acetate.

“Retinoid receptor modulators” refers to compounds which interfere orinhibit the binding of retinoids to the receptor, regardless ofmechanism. Examples of such retinoid receptor modulators includebexarotene, tretinoin, 13-cis-retinoic acid, 9-cis-retinoic acid,α-difluoromethylomithine, ILX23-7553, trans-N-(4′-hydroxyphenyl)retinamide, and N-4-carboxyphenyl retinamide.

“Cytotoxic/cytostatic agents” refer to compounds which cause cell deathor inhibit cell proliferation primarily by interfering directly with thecell's functioning or inhibit or interfere with cell myosis, includingalkylating agents, tumor necrosis factors, intercalators, hypoxiaactivatable compounds, microtubule inhibitors/microtubule-stabilizingagents, inhibitors of mitotic kinesins, inhibitors of kinases involvedin mitotic progression, inhibitors of kinases involved in growth factorand cytokine signal transduction pathways, antimetabolites, biologicalresponse modifiers, hormonal/anti-hormonal therapeutic agents,haematopoietic growth factors, monoclonal antibody targeted therapeuticagents, topoisomerase inhibitors, proteosome inhibitors and ubiquitinligase inhibitors.

Examples of cytotoxic/cytostatic agents include, but are not limited to,sertenef, cachectin, ifosfamide, tasonermin, lonidamine, carboplatin,altretamine, prednimustine, dibromodulcitol, ranimustine, fotemustine,nedaplatin, oxaliplatin, temozolomide, heptaplatin, estramustine,improsulfan tosilate, trofosfamide, nimustine, dibrospidium chloride,pumitepa, lobaplatin, satraplatin, profiromycin, cisplatin, irofulven,dexifosfamide, cis-aminedichloro(2-methyl-pyridine)platinum,benzylguanine, glufosfamide, GPX100, (trans, trans,trans)-bis-mu-(hexane-1,6-diamine)-mu-[diamine-platinum(II)]bis[diamine(chloro)platinum(II)]tetrachloride, diarizidinylspermine, arsenic trioxide,1-(11-dodecylamino-10-hydroxyundecyl)-3,7-dimethylxanthine, zorubicin,idarubicin, daunorubicin, bisantrene, mitoxantrone, pirarubicin,pinafide, valrubicin, amrubicin, antineoplaston,3′-deamino-3′-morpholino-13-deoxo-10-hydroxycaminomycin, annamycin,galarubicin, elinafide, MEN10755,4-demethoxy-3-deamino-3-aziridinyl-4-methylsulphonyl-daunorubicin (seeWO 00/50032), Raf kinase inhibitors (such as Bay43-9006) and mTORinhibitors (such as Wyeth's CCl-779).

An example of a hypoxia activatable compound is tirapazamine

Examples of proteosome inhibitors include but are not limited tolactacystin and MLN-341 (Velcade).

Examples of microtubule inhibitors/microtubule-stabilising agentsinclude paclitaxel, vindesine sulfate,3′,4′-didehydro-4′-deoxy-8′-norvincaleukoblastine, docetaxol, rhizoxin,dolastatin, mivobulin isethionate, auristatin, cemadotin, RPR109881,BMS184476, vinflunine, cryptophycin,2,3,4,5,6-pentafluoro-N-(3-fluoro-4-methoxyphenyl)benzene sulfonamide,anhydrovinblastine,N,N-dimethyl-L-valyl-L-valyl-N-methyl-L-valyl-L-prolyl-L-proline-t-butylamide,TDX258, the epothilones (see for example U.S. Pat. Nos. 6,284,781 and6,288,237) and BMS188797. In an embodiment the epothilones are notincluded in the microtubule inhibitors/microtubule-stabilising agents.

Some examples of topoisomerase inhibitors are topotecan, hycaptamine,irinotecan, rubitecan,6-ethoxypropionyl-3′,4′-O-exo-benzylidene-chartreusin,9-methoxy-N,N-dimethyl-5-nitropyrazolo[3,4,5-k1]acridine-2-(6H)propanamine,1-amino-9-ethyl-5-fluoro-2,3-dihydro-9-hydroxy-4-methyl-1H,12H-benzo[de]pyrano[3′,4′:b,7]-indolizino[1,2b]quinoline-10,13(9H,15H)dione,lurtotecan, 7-[2-(N-isopropylamino)ethyl]-(20S)camptothecin, BNP1350,BNPI1100, BN80915, BN80942, etoposide phosphate, teniposide, sobuzoxane,2′-dimethylamino-2′-deoxy-etoposide, GL331,N-[2-(dimethylamino)ethyl]-9-hydroxy-5,6-dimethyl-6H-pyrido[4,3-b]carbazole-1-carboxamide,asulacrine, (5a, 5aB,8aa,9b)-9-[2-[N-[2-(dimethylamino)ethyl]-N-methylamino]ethyl]-5-[4-hydro0xy-3,5-dimethoxyphenyl]-5,5a,6,8,8a,9-hexohydrofuro(3′,′:6,7)naphtho(2,3-d)-1,3-dioxol-6-one,2,3-(methylenedioxy)-5-methyl-7-hydroxy-8-methoxybenzo[c]-phenanthridinium,6,9-bis[(2-aminoethyl)amino]benzo[g]isoquinoline-5,10-dione,5-(3-aminopropylamino)-7,10-dihydroxy-2-(2-hydroxyethylaminomethyl)-6H-pyrazolo[4,5,1-de]acridin-6-one,N-[1-[2(diethylamino)ethylamino]-7-methoxy-9-oxo-9H-thioxanthen-4-ylmethyl]formamide,N-(2-(dimethylamino)ethyl)acridine-4-carboxamide,6-[[2-(dimethylamino)ethyl]amino]-3-hydroxy-7H-indeno[2,1-c]quinolin-7-one,and dimesna.

Examples of inhibitors of mitotic kinesins, and in particular the humanmitotic kinesin KSP, are described in PCT Publications WO 01/30768 andWO 01/98278, and pending U.S. Ser. Nos. 60/338,779 (filed Dec. 6, 2001),60/338,344 (filed Dec. 6, 2001), 60/338,383 (filed Dec. 6, 2001),60/338,380 (filed Dec. 6, 2001), 60/338,379 (filed Dec. 6, 2001) and60/344,453 (filed Nov. 7, 2001). In an embodiment inhibitors of mitotickinesins include, but are not limited to inhibitors of KSP, inhibitorsof MKLP1, inhibitors of CENP-E, inhibitors of MCAK and inhibitors ofRab6-KIFL.

“Inhibitors of kinases involved in mitotic progression” include, but arenot limited to, inhibitors of aurora kinase, inhibitors of Polo-likekinases (PLK; in particular inhibitors of PLK-1), inhibitors of bub-1and inhibitors of bub-R1.

“Antiproliferative agents” includes antisense RNA and DNAoligonucleotides such as G3139, ODN698, RVASKRAS, GEM231, and INX3001,and antimetabolites such as enocitabine, carmofur, tegafur, pentostatin,doxifluridine, trimetrexate, fludarabine, capecitabine, galocitabine,cytarabine ocfosfate, fosteabine sodium hydrate, raltitrexed,paltitrexid, emitefur, tiazofurin, decitabine, nolatrexed, pemetrexed,nelzarabine, 2′-deoxy-2′-methylidenecytidine,2′-fluoromethylene-2′-deoxycytidine,N-[5-(2,3-dihydro-benzofuryl)sulfonyl]-N′-(3,4-dichlorophenyBurea,N6-[4-deoxy-4-[N2-[2(E),4(E)-tetradecadienoyl]glycylamino]-L-glycero-B-L-manno-heptopyranosyl]adenine,aplidine, ecteinascidin, troxacitabine,4-[2-amino-4-oxo-4,6,7,8-tetrahydro-3H-pyrimidino[5,4-b][1,4]thiazin-6-yl-(S)-ethyl]-2,5-thienoyl-L-glutamicacid, aminopterin, 5-fluorouracil, alanosine,11-acetyl-8-(carbamoyloxymethyl)-4-formyl-6-methoxy-14-oxa-1,11-diazatetracyclo(7.4.1.0.0)-tetradeca-2,4,6-trien-9-yl acetic acid ester, swainsonine,lometrexol, dexrazoxane, methioninase,2′-cyano-2′-deoxy-N4-palmitoyl-1-B-D-arabino furanosyl cytosine,3-aminopyridine-2-carboxaldehyde thiosemicarbazone and trastuzumab.

Examples of monoclonal antibody targeted therapeutic agents includethose therapeutic agents which have cytotoxic agents or radioisotopesattached to a cancer cell specific or target cell specific monoclonalantibody. Examples include Bexxar.

“HMG-CoA reductase inhibitors” refers to inhibitors of3-hydroxy-3-methylglutaryl-CoA reductase. Examples of HMG-CoA reductaseinhibitors that may be used include but are not limited to lovastatin(MEVACOR®; see U.S. Pat. Nos. 4,231,938, 4,294,926 and 4,319,039),simvastatin (ZOCOR®; see U.S. Pat. Nos. 4,444,784, 4,820,850 and4,916,239), pravastatin (PRAVACHOL®; see U.S. Pat. Nos. 4,346,227,4,537,859, 4,410,629, 5,030,447 and 5,180,589), fluvastatin (LESCOL®;see U.S. Pat. Nos. 5,354,772, 4,911,165, 4,929,437, 5,189,164,5,118,853, 5,290,946 and 5,356,896), atorvastatin (LIPITOR®; see U.S.Pat. Nos. 5,273,995, 4,681,893, 5,489,691 and 5,342,952) andcerivastatin (also known as rivastatin and BAYCHOL®; see U.S. Pat. No.5,177,080). The structural formulas of these and additional HMG-CoAreductase inhibitors that may be used in the instant methods aredescribed at page 87 of M. Yalpani, “Cholesterol Lowering Drugs”,Chemistry & Industry, pp. 85-89 (5 Feb. 1996) and U.S. Pat. Nos.4,782,084 and 4,885,314. The term HMG-CoA reductase inhibitor as usedherein includes all pharmaceutically acceptable lactone and open-acidforms (i.e., where the lactone ring is opened to form the free acid) aswell as salt and ester forms of compounds which have HMG-CoA reductaseinhibitory activity, and therefor the use of such salts, esters,open-acid and lactone forms is included within the scope of thisinvention.

“Prenyl-protein transferase inhibitor” refers to a compound whichinhibits any one or any combination of the prenyl-protein transferaseenzymes, including farnesyl-protein transferase (FPTase),geranylgeranyl-protein transferase type I (GGPTase-I), andgeranylgeranyl-protein transferase type-II (GGPTase-II, also called RabGGPTase).

Examples of prenyl-protein transferase inhibitors can be found in thefollowing publications and patents: WO 96/30343, WO 97/18813, WO97/21701, WO 97/23478, WO 97/38665, WO 98/28980, WO 98/29119, WO95/32987, U.S. Pat. No. 5,420,245, U.S. Pat. No. 5,523,430, U.S. Pat.No. 5,532,359, U.S. Pat. No. 5,510,510, U.S. Pat. No. 5,589,485, U.S.Pat. No. 5,602,098, European Patent Publ. 0 618 221, European PatentPubl. 0 675 112, European Patent Publ. 0 604 181, European Patent Publ.0 696 593, WO 94/19357, WO 95/08542, WO 95/11917, WO 95/12612, WO95/12572, WO 95/10514, U.S. Pat. No. 5,661,152, WO 95/10515, WO95/10516, WO 95/24612, WO 95/34535, WO 95/25086, WO 96/05529, WO96/06138, WO 96/06193, WO 96/16443, WO 96/21701, WO 96/21456, WO96/22278, WO 96/24611, WO 96/24612, WO 96/05168, WO 96/05169, WO96/00736, U.S. Pat. No. 5,571,792, WO 96/17861, WO 96/33159, WO96/34850, WO 96/34851, WO 96/30017, WO 96/30018, WO 96/30362, WO96/30363, WO 96/31111, WO 96/31477, WO 96/31478, WO 96/31501, WO97/00252, WO 97/03047, WO 97/03050, WO 97/04785, WO 97/02920, WO97/17070, WO 97/23478, WO 97/26246, WO 97/30053, WO 97/44350, WO98/02436, and U.S. Pat. No. 5,532,359. For an example of the role of aprenyl-protein transferase inhibitor on angiogenesis see European J. ofCancer, Vol. 35, No. 9, pp. 1394-1401 (1999).

“Angiogenesis inhibitors” refers to compounds that inhibit the formationof new blood vessels, regardless of mechanism. Examples of angiogenesisinhibitors include, but are not limited to, tyrosine kinase inhibitors,such as inhibitors of the tyrosine kinase receptors Flt-1 (VEGFR1) andFlk-1/KDR (VEGFR2), inhibitors of epidermal-derived, fibroblast-derived,or platelet derived growth factors, MMP (matrix metalloprotease)inhibitors, integrin blockers, interferon-α, interleukin-12, pentosanpolysulfate, cyclooxygenase inhibitors, including nonsteroidalanti-inflammatories (NSAIDs) like aspirin and ibuprofen as well asselective cyclooxy-genase-2 inhibitors like celecoxib and rofecoxib(PNAS, Vol. 89, p. 7384 (1992); JNCI, Vol. 69, p. 475 (1982); Arch.Opthalmol., Vol. 108, p. 573 (1990); Anat. Rec., Vol. 238, p. 68 (1994);FEBS Letters, Vol. 372, p. 83 (1995); Clin, Orthop. Vol. 313, p. 76(1995); J. Mol. Endocrinol., Vol. 16, p. 107 (1996); Jpn. J. Pharmacol.,Vol. 75, p. 105 (1997); Cancer Res., Vol. 57, p. 1625 (1997); Cell, Vol.93, p. 705 (1998); Intl. J. Mol. Med., Vol. 2, p. 715 (1998); J. Biol.Chem., Vol. 274, p. 9116 (1999)), steroidal anti-inflammatories (such ascorticosteroids, mineralocorticoids, dexamethasone, prednisone,prednisolone, methylpred, betamethasone), carboxyamidotriazole,combretastatin A-4, squalamine, 6-O-chloroacetyl-carbonyl)-fumagillol,thalidomide, angiostatin, troponin-1, angiotensin II antagonists (seeFernandez et al., J. Lab. Clin. Med. 105:141-145 (1985)), and antibodiesto VEGF (see, Nature Biotechnology, Vol. 17, pp. 963-968 (October 1999);Kim et al., Nature, 362, 841-844 (1993); WO 00/44777; and WO 00/61186).

Other therapeutic agents that modulate or inhibit angiogenesis and mayalso be used in combination with the compounds of the instant inventioninclude agents that modulate or inhibit the coagulation and fibrinolysissystems (see review in Clin. Chem. La. Med. 38:679-692 (2000)). Examplesof such agents that modulate or inhibit the coagulation and fibrinolysispathways include, but are not limited to, heparin (see Thromb. Haemost.80:10-23 (1998)), low molecular weight heparins and carboxypeptidase Uinhibitors (also known as inhibitors of active thrombin activatablefibrinolysis inhibitor [TAFIa]) (see Thrombosis Res. 101:329-354(2001)). TAFIa inhibitors have been described in U.S. Ser. Nos.60/310,927 (filed Aug. 8, 2001) and 60/349,925 (filed Jan. 18, 2002).

“Agents that interfere with cell cycle checkpoints” refer to compoundsthat inhibit protein kinases that transduce cell cycle checkpointsignals, thereby sensitizing the cancer cell to DNA damaging agents.Such agents include inhibitors of ATR, ATM, the Chk1 and Chk2 kinasesand cdk and cdc kinase inhibitors and are specifically exemplified by7-hydroxystaurosporin, flavopiridol, CYC202 (Cyclacel) and BMS-387032.

“Inhibitors of cell proliferation and survival signalling pathway” referto compounds that inhibit signal transduction cascades downstream ofcell surface receptors. Such agents include inhibitors ofserine/threonine kinases (including but not limited to inhibitors of Aktsuch as described in WO 02/083064, WO 02/083139, WO 02/083140 and WO02/083138), inhibitors of Raf kinase (for example BAY-43-9006),inhibitors of MEK (for example CI-1040 and PD-098059), inhibitors ofmTOR (for example Wyeth CCl-779), and inhibitors of PI3K (for exampleLY294002).

As described above, the combinations with NSAID's are directed to theuse of NSAID's which are potent COX-2 inhibiting agents. For purposes ofthis specification an NSAID is potent if it possesses an IC₅₀ for theinhibition of COX-2 of 1 μM or less as measured by cell or microsomalassays.

The invention also encompasses combinations with NSAID's which areselective COX-2 inhibitors. For purposes of this specification NSAID'swhich are selective inhibitors of COX-2 are defined as those whichpossess a specificity for inhibiting COX-2 over COX-1 of at least 100fold as measured by the ratio of IC₅₀ for COX-2 over IC₅₀ for COX-1evaluated by cell or microsomal assays. Such compounds include, but arenot limited to those disclosed in U.S. Pat. No. 5,474,995, U.S. Pat. No.5,861,419, U.S. Pat. No. 6,001,843, U.S. Pat. No. 6,020,343, U.S. Pat.No. 5,409,944, U.S. Pat. No. 5,436,265,

U.S. Pat. No. 5,536,752, U.S. Pat. No. 5,550,142, U.S. Pat. No.5,604,260, U.S. Pat. No. 5,698,584, U.S. Pat. No. 5,710,140, WO94/15932, U.S. Pat. No. 5,344,991, U.S. Pat. No. 5,134,142, U.S. Pat.No. 5,380,738, U.S. Pat. No. 5,393,790, U.S. Pat. No. 5,466,823, U.S.Pat. No. 5,633,272 and U.S. Pat. No. 5,932,598, all of which are herebyincorporated by reference.

Inhibitors of COX-2 that are particularly useful in the instant methodof treatment are: 3-phenyl-4-(4-(methylsulfonyl)phenyl)-2-(5H)-furanone;and5-chloro-3-(4-methylsulfonyl)phenyl-2-(2-methyl-5-pyridinyl)pyridine; ora pharmaceutically acceptable salt thereof.

Compounds that have been described as specific inhibitors of COX-2 andare therefore useful in the present invention include, but are notlimited to, the following: parecoxib, BEXTRA® and CELEBREX® or apharmaceutically acceptable salt thereof.

Other examples of angiogenesis inhibitors include, but are not limitedto, endostatin, ukrain, ranpirnase, IM862,5-methoxy-4-[2-methyl-3-(3-methyl-2-butenyl)oxiranyl]-1-oxaspiro[2,5]oct-6-yl(chloroacetyl)carbamate,acetyldinanaline,5-amino-1-[[3,5-dichloro-4-(4-chlorobenzoyl)phenyl]methyl]-1H-1,2,3-triazole-4-carboxamide,CM101, squalamine, combretastatin, RPI4610, NX31838, sulfatedmannopentaose phosphate,7,7-(carbonyl-bis[imino-N-methyl-4,2-pyrrolocarbonylimino[N-methyl-4,2-pyrrole]-carbonylimino]-bis-(1,3-naphthalenedisulfonate), and 3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone(SU5416).

As used above, “integrin blockers” refers to compounds which selectivelyantagonize, inhibit or counteract binding of a physiological ligand tothe α_(v)β₃ integrin, to compounds which selectively antagonize, inhibitor counteract binding of a physiological ligand to the αvβ5 integrin, tocompounds which antagonize, inhibit or counteract binding of aphysiological ligand to both the α_(v)β₃ integrin and the α_(v)β₅integrin, and to compounds which antagonize, inhibit or counteract theactivity of the particular integrin(s) expressed on capillaryendothelial cells. The term also refers to antagonists of the α_(v)β₆,α_(v)β₈, α₁β₁, α₂β₁, α₅β₁, α₆β₁ and α₆β₄ integrins. The term also refersto antagonists of any combination of α_(v)β₃, α_(v)β₅, α_(v)β₆, α_(v)β₈,a₁β₁, α₂β₁, α₅β₁, α₆β₁ and α₆β₄ integrins.

Some specific examples of tyrosine kinase inhibitors includeN-(trifluoromethylphenyl)-5-methylisoxazol-4-carboxamide,3-[(2,4-dimethylpyrrol-5-yl)methylidenyl)indolin-2-one,17-(allylamino)-17-demethoxygeldanamycin,4-(3-chloro-4-fluorophenylamino)-7-methoxy-6-[3-(4-morpholinyl)propoxyl]quinazoline,N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine,BIBX1382,2,3,9,10,11,12-hexahydro-10-(hydroxymethyl)-10-hydroxy-9-methyl-9,12-epoxy-1H-diindolo[1,2,3-fg:3′,2′,1′-k1]pyrrolo[3,4-i][1,6]benzodiazocin-1-one,SH268, genistein, STI5711, CEP2563,4-(3-chlorophenylamino)-5,6-dimethyl-7H-pyrrolo[2,3-d]pyrimidinemethanesulfonate, 4-(3-bromo-4-hydroxyphenyl)amino-6,7-dimethoxyquinazoline,4-(4′-hydroxyphenyl)amino-6,7-dimethoxyquinazoline, SU6668, STI571A,N-4-chlorophenyl-4-(4-pyridylmethyl)-1-phthalazinamine, and EMD121974.

Combinations with compounds other than anti-cancer compounds are alsoencompassed in the instant methods. For example, combinations of theinstantly claimed compounds with PPAR-γ (i.e., PPAR-gamma) agonists andPPAR-δ (i.e., PPAR-delta) agonists are useful in the treatment ofcertain malingnancies. PPAR-γ and PPAR-δ are the nuclear peroxisomeproliferator-activated receptors γ and δ. The expression of PPAR-γ onendothelial cells and its involvement in angiogenesis has been reportedin the literature (see J. Cardiovasc. Pharmacol. 1998; 31:909-913; J.Biol. Chem. 1999; 274:9116-9121; Invest. Opthalmol. Vis. Sci. 2000;41:2309-2317). More recently, PPAR-γ agonists have been shown to inhibitthe angiogenic response to VEGF in vitro; both troglitazone androsiglitazone maleate inhibit the development of retinalneovascularization in mice. (Arch. Ophthamol. 2001; 119:709-717).Examples of PPAR-γ agonists and PPAR-γ/α agonists include, but are notlimited to, thiazolidinediones (such as DRF2725, CS-011, troglitazone,rosiglitazone, and pioglitazone), fenofibrate, gemfibrozil, clofibrate,GW2570, SB219994, AR-H039242, ITT-501, MCC-555, GW2331, GW409544,NN2344, KRP297, NP0110, DRF4158, NN622, GI262570, PNU182716, DRF552926,2-[(5,7-dipropyl-3-trifluoromethyl-1,2-benzisoxazol-6-yl)oxy]-2-methylpropionicacid (disclosed in U.S. Ser. No. 09/782,856), and2(R)-7-(3-(2-chloro-4-(4-fluorophenoxy)phenoxy)propoxy)-2-ethylchromane-2-carboxylicacid (disclosed in U.S. Ser. No. 60/235,708 and 60/244,697).

Another embodiment of the instant invention is the use of the presentlydisclosed compounds in combination with gene therapy for the treatmentof cancer. For an overview of genetic strategies to treating cancer seeHall et al (Am. J. Hum. Genet. 61:785-789, 1997) and Kufe et al (CancerMedicine, 5th Ed, pp 876-889, B C Decker, Hamilton 2000). Gene therapycan be used to deliver any tumor suppressing gene. Examples of suchgenes include, but are not limited to, p53, which can be delivered viarecombinant virus-mediated gene transfer (see U.S. Pat. No. 6,069,134,for example), a uPA/uPAR antagonist (“Adenovirus-Mediated Delivery of auPA/uPAR Antagonist Suppresses Angiogenesis-Dependent Tumor Growth andDissemination in Mice,” Gene Therapy, August 1998; 5(8):1105-13), andinterferon gamma (J. Immunol. 2000; 164:217-222).

The compounds of the instant invention may also be administered incombination with an inhibitor of inherent multidrug resistance (MDR), inparticular MDR associated with high levels of expression of transporterproteins. Such MDR inhibitors include inhibitors of p-glycoprotein(P-gp), such as LY335979, XR9576, OC144-093, R101922, VX853 and PSC833(valspodar).

A compound of the present invention may be employed in conjunction withanti-emetic agents to treat nausea or emesis, including acute, delayed,late-phase, and anticipatory emesis, which may result from the use of acompound of the present invention, alone or with radiation therapy. Forthe prevention or treatment of emesis, a compound of the presentinvention may be used in conjunction with other anti-emetic agents,especially neurokinin-1 receptor antagonists, 5HT3 receptor antagonists,such as ondansetron, granisetron, tropisetron, and zatisetron, GABABreceptor agonists, such as baclofen, a corticosteroid such as Decadron(dexamethasone), Kenalog, Aristocort, Nasalide, Preferid, Benecorten orothers such as disclosed in U.S. Pat. Nos. 2,789,118, 2,990,401,3,048,581, 3,126,375, 3,929,768, 3,996,359, 3,928,326 and 3,749,712, anantidopaminergic, such as the phenothiazines (for exampleprochlorperazine, fluphenazine, thioridazine and mesoridazine),metoclopramide or dronabinol. In another embodiment, conjunctive therapywith an anti-emesis agent selected from a neurokinin-1 receptorantagonist, a 5HT3 receptor antagonist and a corticosteroid is disclosedfor the treatment or prevention of emesis that may result uponadministration of the instant compounds.

Neurokinin-1 receptor antagonists of use in conjunction with thecompounds of the present invention are fully described, for example, inU.S. Pat. Nos. 5,162,339, 5,232,929, 5,242,930, 5,373,003, 5,387,595,5,459,270, 5,494,926, 5,496,833, 5,637,699, 5,719,147; European PatentPublication Nos. EP 0 360 390, 0 394 989, 0 428 434, 0 429 366, 0 430771, 0 436 334, 0 443 132, 0 482 539, 0 498 069, 0 499 313, 0 512 901, 0512 902, 0 514 273, 0 514 274, 0 514 275, 0 514 276, 0 515 681, 0 517589, 0 520 555, 0 522 808, 0 528 495, 0 532 456, 0 533 280, 0 536 817, 0545 478, 0 558 156, 0 577 394, 0 585 913, 0 590 152, 0 599 538, 0 610793, 0 634 402, 0 686 629, 0 693 489, 0 694 535, 0 699 655, 0 699 674, 0707 006, 0 708 101, 0 709 375, 0 709 376, 0 714 891, 0 723 959, 0 733632 and 0 776 893; PCT International Patent Publication Nos. WO90/05525, 90/05729, 91/09844, 91/18899, 92/01688, 92/06079, 92/12151,92/15585, 92/17449, 92/20661, 92/20676, 92/21677, 92/22569, 93/00330,93/00331, 93/01159, 93/01165, 93/01169, 93/01170, 93/06099, 93/09116,93/10073, 93/14084, 93/14113, 93/18023, 93/19064, 93/21155, 93/21181,93/23380, 93/24465, 94/00440, 94/01402, 94/02461, 94/02595, 94/03429,94/03445, 94/04494, 94/04496, 94/05625, 94/07843, 94/08997, 94/10165,94/10167, 94/10168, 94/10170, 94/11368, 94/13639, 94/13663, 94/14767,94/15903, 94/19320, 94/19323, 94/20500, 94/26735, 94/26740, 94/29309,95/02595, 95/04040, 95/04042, 95/06645, 95/07886, 95/07908, 95/08549,95/11880, 95/14017, 95/15311, 95/16679, 95/17382, 95/18124, 95/18129,95/19344, 95/20575, 95/21819, 95/22525, 95/23798, 95/26338, 95/28418,95/30674, 95/30687, 95/33744, 96/05181, 96/05193, 96/05203, 96/06094,96/07649, 96/10562, 96/16939, 96/18643, 96/20197, 96/21661, 96/29304,96/29317, 96/29326, 96/29328, 96/31214, 96/32385, 96/37489, 97/01553,97/01554, 97/03066, 97/08144, 97/14671, 97/17362, 97/18206, 97/19084,97/19942 and 97/21702; and in British Patent Publication Nos. 2 266 529,2 268 931, 2 269 170, 2 269 590, 2 271 774, 2 292 144, 2 293 168, 2 293169, and 2 302 689. The preparation of such compounds is fully describedin the aforementioned patents and publications, which are incorporatedherein by reference.

In an embodiment, the neurokinin-1 receptor antagonist for use inconjunction with the compounds of the present invention is selectedfrom:2-(R)-(1-(R)-(3,5-bis(trifluoromethyl)phenyl)ethoxy)-3-(S)-(4-fluorophenyl)-4-(3-(5-oxo-1H,4H-1,2,4-triazolo)methyl)morpholine,or a pharmaceutically acceptable salt thereof, which is described inU.S. Pat. No. 5,719,147.

A compound of the instant invention may also be administered with anagent useful in the treatment of anemia. Such an anemia treatment agentis, for example, a continuous eythropoiesis receptor activator (such asepoetin alfa).

A compound of the instant invention may also be administered with anagent useful in the treatment of neutropenia. Such a neutropeniatreatment agent is, for example, a hematopoietic growth factor whichregulates the production and function of neutrophils such as a humangranulocyte colony stimulating factor, (G-CSF). Examples of a G-CSFinclude filgrastim.

A compound of the instant invention may also be administered with animmunologic-enhancing drug, such as levamisole, isoprinosine andZadaxin.

Thus, the scope of the instant invention encompasses the use of theinstantly claimed compounds in combination with a second compoundselected from: an estrogen receptor modulator, an androgen receptormodulator, a retinoid receptor modulator, a cytotoxic/cytostatic agent,an antiproliferative agent, a prenyl-protein transferase inhibitor, anHMG-CoA reductase inhibitor, an HIV protease inhibitor, a reversetranscriptase inhibitor, an angiogenesis inhibitor, PPAR-γ agonists,PPAR-δ agonists, an inhibitor of inherent multidrug resistance, ananti-emetic agent, an agent useful in the treatment of anemia, an agentuseful in the treatment of neutropenia, an immunologic-enhancing drug,an inhibitor of cell proliferation and survival signaling, and an agentthat interferes with a cell cycle checkpoint.

The term “administration” and variants thereof (e.g., “administering” acompound) in reference to a compound of the invention means introducingthe compound or a prodrug of the compound into the system of the animalin need of treatment. When a compound of the invention or prodrugthereof is provided in combination with one or more other active agents(e.g., a cytotoxic agent, etc.), “administration” and its variants areeach understood to include concurrent and sequential introduction of thecompound or prodrug thereof and other agents.

As used herein, the term “composition” is intended to encompass aproduct comprising the specified ingredients in the specified amounts,as well as any product which results, directly or indirectly, fromcombination of the specified ingredients in the specified amounts.

The term “therapeutically effective amount” as used herein means thatamount of active compound or pharmaceutical agent that elicits thebiological or medicinal response in a tissue, system, animal or humanthat is being sought by a researcher, veterinarian, medical doctor orother clinician.

The term “treating cancer” or “treatment of cancer” refers toadministration to a mammal afflicted with a cancerous condition andrefers to an effect that alleviates the cancerous condition by killingthe cancerous cells, but also to an effect that results in theinhibition of growth and/or metastasis of the cancer.

In an embodiment, the angiogenesis inhibitor to be used as the secondcompound is selected from a tyrosine kinase inhibitor, an inhibitor ofepidermal-derived growth factor, an inhibitor of fibroblast-derivedgrowth factor, an inhibitor of platelet derived growth factor, an MMP(matrix metalloprotease) inhibitor, an integrin blocker, interferon-α,interleukin-12, pentosan polysulfate, a cyclooxygenase inhibitor,carboxyamidotriazole, combretastatin A-4, squalamine,6-O-chloroacetyl-carbonyl)-fumagillol, thalidomide, angiostatin,troponin-1, or an antibody to VEGF. In an embodiment, the estrogenreceptor modulator is tamoxifen or raloxifene.

Also included in the scope of the claims is a method of treating cancerthat comprises administering a therapeutically effective amount of acompound of Formula A in combination with radiation therapy and/or incombination with a second compound selected from: an estrogen receptormodulator, an androgen receptor modulator, a retinoid receptormodulator, a cytotoxiccytostatic agent, an antiproliferative agent, aprenyl-protein transferase inhibitor, an HMG-CoA reductase inhibitor, anHIV protease inhibitor, a reverse transcriptase inhibitor, anangiogenesis inhibitor, PPAR-γ agonists, PPAR-δ agonists, an inhibitorof inherent multidrug resistance, an anti-emetic agent, an agent usefulin the treatment of anemia, an agent useful in the treatment ofneutropenia, an immunologic-enhancing drug, an inhibitor of cellproliferation and survival signaling, and an agent that interferes witha cell cycle checkpoint.

And yet another embodiment of the invention is a method of treatingcancer that comprises administering a therapeutically effective amountof a compound of Formula A in combination with paclitaxel ortrastuzumab.

The invention further encompasses a method of treating or preventingcancer that comprises administering a therapeutically effective amountof a compound of Formula A in combination with a COX-2 inhibitor.

The instant invention also includes a pharmaceutical composition usefulfor treating or preventing cancer that comprises a therapeuticallyeffective amount of a compound of Formula A and a second compoundselected from: an estrogen receptor modulator, an androgen receptormodulator, a retinoid receptor modulator, a cytotoxic/cytostatic agent,an antiproliferative agent, a prenyl-protein transferase inhibitor, anHMG-CoA reductase inhibitor, an HIV protease inhibitor, a reversetranscriptase inhibitor, an angiogenesis inhibitor, a PPAR-γ agonist, aPPAR-δ agonist, an inhibitor of cell proliferation and survivalsignaling, and an agent that interferes with a cell cycle checkpoint.

All patents, publications and pending patent applications identified arehereby incorporated by reference.

Abbreviations used in the description of the chemistry and in theExamples that follow are: AEBSF (p-aminoethylbenzenesulfonyl fluoride);BSA (bovine serum albumin); BuLi (n-Butyl lithium); CDCl₃(chloroform-d); CuI (copper iodide); CuSO₄ (copper sulfate); DCE(dichloroethane); DCM (dichloromethane); DEAD (diethylazodicarboxylate); DMF (N,N-dimethylformamide); DMSO (dimethylsulfoxide); DTT (dithiothreitol); EDTA (ethylene-diamine-tetra-aceticacid); EGTA (ethylene-glycol-tetra-acetic acid); EtOAc (ethyl acetate);EtOH (ethanol); HOAc (acetic acid); HPLC (high-performance liquidchromatography); HRMS (high resolution mass spectrum); LCMS (liquidchromatograph-mass spectrometer); LHMDS (lithiumbis(trimethylsilyl)amide); LRMS (low resolution mass spectrum); MeOH(methanol); MP-B(CN)H₃ (Macroporous cyanoborohydride); NaHCO₃ (sodiumbicarbonate); Na₂SO₄ (sodium sulfate); Na(OAc)₃BH (sodiumtriacetoxyborohydride); NH₄OAc (ammonium acetate); NBS(N-bromosuccinamide); NMR (nuclear magnetic resonance); PBS (phosphatebuffered saline); PCR (polymerase chain reaction); Pd(dppf)([1,1′-bis(diphenylphosphino)ferrocene]palladium); Pd(Ph₃)₄(palladium(0)tetrakis-triphenylphosphine); POCl₃ (phosphorous oxychloride); PS-DIEA(polystyrene diisopropylethylamine); TBAF (tetrabutylammonium fluoride);THF (tetrahydrofuran); TFA (trifluoroacteic acid); and TMSCH₂N₂(trimethylsilyldiazomethane).

The compounds of this invention may be prepared by employing reactionsas shown in the following Reaction Schemes, in addition to otherstandard manipulations that are known in the literature or exemplifiedin the experimental procedures. The illustrative Reaction Schemes below,therefore, are not limited by the compounds listed or by any particularsubstituents employed for illustrative purposes. Substituent numberingas shown in the Reaction Schemes does not necessarily correlate to thatused in the claims and often, for clarity, a single substituent is shownattached to the compound where multiple substituents are optionallyallowed under the definitions of Formula A hereinabove.

Reactions used to generate the compounds of this invention are preparedby employing reactions as shown in the Reaction Schemes I-III, inaddition to other standard manipulations such as ester hydrolysis,cleavage of protecting groups, etc., as may be known in the literatureor exemplified in the experimental procedures.

Synopsis of Reaction Schemes

Reaction Scheme I illustrates the preparation of the pyridazinecompounds of the instant invention, starting with4-phenyl-3,6-dichloropyridazine I-1. This material can be reacted with anucleophile to give I-2. When the nucleophile is an amine, heating undermicrowave conditions is preferred and when the nucleophile is analcohol, the metal alkoxide is used. In addition, a (hetero)aryl groupcan be introduced under Suzuki coupling conditions. In all cases thesubstitution occurs predominately at the less hindered 6-position.Intermediate I-2 can be subjected to a standard Suzuki coupling with(4-formylphenyl) boronic acid to afford I-3. This material can thenundergo reductive amination with a diverse array of amines, for example,

and a borohydride to provide I-4.

Reaction Scheme II illustrates the synthesis of the pyrimidine compoundsof the instant invention, starting with methyl(2Z)-3-methoxy-3-phenylacrylate II-1. Condensation of II-1 with amidineor guanidines provides II-2, which reacts with POCl₃ to affordpyrimidine II-3. Suzuki coupling of II-3 with (4-formylphenyl)boronicacid produces the aldehyde II-4. Reductive amination with suitablysubstituted amines generates amines II-5.

Reaction Scheme III illustrates the preparation of the4-(5-phenylpyrimidin-4-yl)benzyl compounds. Synthesis starts with acommercially available carboxaldehyde (III-1), which is converted to thecorresponding acetal and then treated with a Grignard reagent to produceketone (III-2). Ketone (III-2) is then formylated using a formyl groupdonor such as dimethylformamide-dimethyl acetal to produce enamine(III-3). The enamine can then be converted to the thiopyrimidine by thetreatment with thiourea and a base such as potassiumt-butoxide, sodiummethoxide or sodium hydroxide in a solvent such as methanol or ethanol.The thiopyrimidine is then alkylated in situ using an alkylating reagentsuch as methyl iodide or benzyl bromide to produce thealkylthiopyrimidine (III-4). Oxidation to the sulfone can be easilyachieved by those skilled in the art. One possible method is a two stepprocedure using peracetic acid to produce the sulfoxide and thenhydrogen peroxide and catalytic sodium tungstenate to provide sulfone(III-5). Sulfone (III-5) is a versatile intermediate and can be treatedwith a variety of amines in dioxane with or without a base such assodium bis(trimethylsilyl)amide in solvents including, but not limitedto dioxane or THF with or without heating in a microwave. The acetal isthen removed under acidic conditions and the liberated aldehyde can beconverted to the amine using reductive amination conditions to produce(III-6). Alternatively, sulfone (III-5) can be treated with potassiumcyanide in DMSO with or without heating. The acetal could then beremoved under acidic conditions and treatment of the aldehyde with anamine under reductive conditions produces the corresponding alkylatedproduct. Finally, treatment with thiosemicarbazide in TFA producesaminothiotriazole (III-7).

EXAMPLES

Examples provided are intended to assist in a further understanding ofthe invention. Particular materials employed, species and conditions areintended to be further illustrative of the invention and not limitativeof the reasonable scope thereof. The reagents utilized in synthesizingthe compounds depicted in the following Tables are either commerciallyavailable or are readily prepared by one of ordinary skill in the art.

N,N-dimethyl-5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-pyrazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazin-3-amine(1-5)

-   4-Phenyl-3,6-dichloropyridazine (1-1; 226 mg, 1.0 mmol) (prepared as    described in U.S. Pat. No. 6,444,666) was added to a mixture of    diisopropylethylamine (0.16 mL, 1 mmol) and 2M    dimethylamine/methanol (1.0 mL, 2 0 mmol) in n-butanol (1 mL),    sealed in a reaction tube and heated in a microwave for two 15 min.    cycles. The reaction mixture was evaporated to remove solvents and    the residue was dissolved in ethyl acetate, washed with    sat'd.aqueous Na₂CO₃, the solvent dried over anhydrous Na₂ SO₄, and    solution filtered and evaporated to give a mixture of two isomeric    products, the predominant product being the desired product. This    residue was triturated with diethyl ether/hexane to give    N,N-dimethyl 6-chloro-5-phenylpyridazin-3-amine (1-2) as a    crystalline solid (132 mg). Additional material could be obtained by    chromatographic separation of the mother liquors on silica gel by    gradient elution with ethyl acetate/hexane. 1H NMR (CDCl₃): δ7.48    (5H, s), 6.73 (1H, s), 3.20 (6H, s); m/e (m+1): 234.2

N,N-dimethyl 6-chloro-5-phenylpyridazin-3-amine (1-2; 120 mg, 0.51 mmol)was dissolved in dry dioxane (3 mL) and 4-formylphenyl boronic acid (136mg, 0.91 mmol). 2 M aqueous Na₂CO₃ (0.75 mL) and palladium(0)tetrakis-triphenylphosphine (66 mg, 0.058 mmol) were then added. Thismixture was degassed, the vessel sealed and heated at 110° C. for 24hours. The cooled reaction was diluted with water and the productextracted into ethyl acetate, the extract dried over anhydrous NaSO₄,filtered and evaporated. The product was purified by chromatography onsilica gel and elution with a 25-65% ethyl acetate/hexane gradient togive 4-[6-(Dimethylamino)-4-phenylpyridazin-3-yl]benzaldehyde (1-3). NMR(CDCl₃): δ9.98 (1H, s), 7.74 (2H, d, J=8 Hz), 7.54 (2H, d, J=8 Hz), 7.35(3H, m), 7.19 (2H, d, J=8 Hz), 6.76 (1H, s), 3.29 (6H, s); m/e (m+1):304.2

4-[6-(Dimethylamino)-4-phenylpyridazin-3-yl]benzaldehyde (1-3: 76 mg,0.25 mmol) was dissolved in dry dimethylformamide (2 mL). Then2-(3-piperidin-4-yl-1H-pyazol-5-yl)pyridine (1-4 [see synthesis below];62 mg, 0.27 mmol) and acetic acid (0.06 mL, 1.0 mmol) were added. Afterstirring for 15 min., sodium triacetoxyborohydride (120 mg, 0.58 mmol)was added and the reaction mixture was stirred at ambient temperaturefor 12 hours. This reaction mixture was partially evaporated and theresidue chromatographed on a silica gel column eluting with a 0-7%methanol/ethyl acetate (sat'd with NH₄OH).N,N-dimethyl-5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-pyrazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazin-3-amine(1-5) was obtained as a white solid upon trituration with diethylether/hexane. NMR (CDCl₃): δ8.57 (1H, d, J=4.5 Hz), 7.68-7.73 (2H, m),7.28-7.35 (6H, m), 7.20 (4H, br d, J=7 Hz), 6.74 (1H, s), 6.60 (1H, s),3.35 (2H, s), 3.26 (6H, s), 2.96 (2H, d, J=11 Hz), 2.72 (1H, m), 2.10(2H, br t, J=11 Hz), 1.97 (2H, br d, J=12 Hz), 1.81 (2H, br q, J=11 Hz);m/e (m+1): 516.3.

2-(3-Piperidin-4-yl-1H-pyazol-5-yl)pyridine (1-4)

tert-Butyl 4-acetylpiperidine-1-carboxylate (described in U.S. Pat. No.5,952,341, 4.00 g, 17.6 mmol) was dissolved in 40 ml of anhydrous THFunder N2. NaH (60% suspension, 0.93 g, 38.7 mmol) was added followed bythe addition of ethyl pyridine-2-carboxylate (2.38 ml, 17.6 mmol). Thereaction was heated to 60° C. After 2 h anhydrous DMF (20 ml) was addedand heating was continued for 2 h. The reaction was allowed to cool toambient temperature and was quenched with saturated aqueous NH₄Cl. Themixture was extracted 3× with DCM and the combined organic extracts weredried over Na₂SO₄, filtered and concentrated in vacuo. The resulting oilconsisting primarily of tert-butyl4-(3-oxo-3-pyridin-2-ylpropanoyl)piperidine-1-carboxylate was usedwithout purification.

Unpurified tert-butyl4-(3-oxo-3-pyridin-2-ylpropanoyl)piperidine-1-carboxylate (2.00 g, 6.02mmol) was dissolved in 20 ml of EtOH and hydrazine hydrate (0.321 ml,6.62 mmol) was added and the reaction was warmed to 80° C. After 15 hthe reaction was concentrated in vacuo and the residue was purified byflash column chromatography (dissolve in DCM, elute with 98:2 to 95:5DCM/MeOH). tert-Butyl4-(3-pyridin-2-yl-1H-pyrazol-5-yl)piperidine-1-carboxylate was obtainedas a yellow oil. ¹H NMR (CDCl₃): δ 8.60 (m, 1H), 7.73 (dt, J=1.7, 7.6Hz, 1H), 7.65 (d, J=7.8 Hz, 1H), 7.22 (m, 1H), 6.57 (s, 1H), 4.18 (bs,2H), 2.89 (m, 4H), 2.00 (d, J=12.5 Hz, 2H), 1.67 (m, 2H), 1.48 (s, 9H).m/z (m+1): 329.2

tert-Butyl 4-(3-pyridin-2-yl-1H-pyrazol-5-yl)piperidine-1-carboxylate(1.46 g, 4.45 mmol) was dissolved in 15 ml DCM. Trifluoroacetic acid (5ml) was added gradually and the resulting solution was stirred atambient temperature for 3 h and was then concentrated in vacuo. Theresidue was dissolved in 4:1 MeOH/water and treated with MP-carbonateresin (5 g, 2.55 mmol/g). After stirring overnight the reaction wasfiltered and the filtrate was concentrated in vacuo to provide 1.1 g of2-(3-piperidin-4-yl-1H-pyrazol-5-yl)pyridine (1-4) as a tan solid. 68.56 (d, J=3.9 Hz, 1H), 7.84 (m, 2H), 7.21 (m, 1H), 6.65 (s, 1H), 3.22(m, 3H), 2.87 (m, 2H), 2.03 (d, J=12.8 Hz, 2H), 1.71 (d, J=10.3 Hz, 2H).m/z (m+1): 229.3.

The following compounds in Tables 1a and 1b were prepared by theprocedures described in Scheme 1 substituting the appropriate amines atthe first and last steps and where necessary oxidation of the morpholinering or removal of a Boc protecting group.

TABLE 1a Cmpd MS No. Structure Nomenclature M + 1 1-6

N-methyl-5-phenyl-6-(4-{[4-(5- pyridin-2-yl-1H-pyrazol-3-yl)piperidin-1- yl]methyl}phenyl)pyridazin-3-amine 502.2

TABLE 1b

Cmpd Structure MS No. R¹ Nomenclature M + 1 1-7

4-[5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)-piperidin-1-yl]methyl}phenyl)pyridazin-3-yl]morpholine 559.3 1-8

tert-butyl 4-[5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazin-3-yl]piperazine-1-carboxylate 658.6 1-9

4-phenyl-6-piperazin-1-yl-3-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazine 558.1 1-10

4-[5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazin-3-yl]thiomorpholine 575.4 1-11

4-[5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazin-3-yl]thiomorpholine- 1-oxide591.3 1-12

4-[5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazin-3-yl]thiomorpholine-1,1-dioxide 607.3

3-methoxy-5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-pyrazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazine(2-3) and5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-pyrazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazin-3-ol(2-4)

A commercial solution of 25 wt % sodium methoxide in methanol (0.49 mL,2 0 mmol) was added drop wise to an ice-cooled solution of4-phenyl-3,6-dichloropyridazine (1-1; 448 mg, 2.0 mmol) in dry methanol(10 mL). This solution was gradually warmed to room temperature overthree hours. The solvent was evaporated and the residue dissolved inmethylene chloride, washed with aq. NaHCO₄, the organic layer dried overanhydrous Na₂SO₄, filtered and evaporated to give a 2:1 mixture ofisomeric products. Upon chromatography on silica gel and elution with15% ethyl acetate/hexane, the desired and major isomer was isolated.Trituration of this material with diethyl ether/hexane afforded3-chloro-6-methoxy-4-phenylpyridazine as a white solid (2-1). NMR(CDCl₃): δ7.49 (5H, br s), 6.96 (1H, s), 4.17 (3H, s).

3-Chloro-6-methoxy-4-phenylpyridazine (2-1; 179 mg, 0.81 mmol) wasdissolved in dry dioxane (4 mL) and 4-formylphenyl boronic acid (165 mg,1.1 mmol), 2 M aqueous Na₂CO₃ (1.2 mL) and palladium(0)tetrakis-triphenylphosphine (108 mg, 0.09 mmol) were added. This mixturewas degassed, the vessel sealed and heated at 110° C. for 24 hours. Thecooled reaction was diluted with water and the product extracted intoethyl acetate, the extract dried over anhydrous NaSO₄, filtered andevaporated. This residue was chromatographed on silica gel and elutedwith a 25-45% ethyl acetate/hexane gradient to4-(6-methoxy-4-phenylpyridazin-3-yl)benzaldehyde (2-2) as a viscous oil.NMR (CDCl₃): δ10.0 (1H, s), 7.79 (2H, d, J=8 Hz), 7.54 (2H, d, J=8 Hz),7.31-7.37 (3H, m), 7.15 (2H, d, J=8 Hz), 7.03 (1H, s), 4.24 (3H, s).

4-(6-Methoxy-4-phenylpyridazin-3-yl)benzaldehyde (2-2; 80 mg, 0.27 mmol)was dissolved in dry dimethylformamide (2 mL) and2-(3-piperidin-4-yl-1H-pyazol-5-yl)pyridine (1-4; 99 mg, 0.43 mmol) andacetic acid (0.14 mL, 2.3 mmol) were added. After stirring for 15 min.,sodium triacetoxyborohydride (116 mg, 0.54 mmol) was added and thereaction mixture was stirred at ambient temperature for 12 hours. Thisreaction mixture was partially evaporated and the residuechromatographed on a silica gel column eluting with a 0-5%methanol/ethyl acetate sat'd with NH₄OH.3-methoxy-5-phenyl-6-(4-{[445-pyridin-2-yl-1H-pyrazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazine(2-3) was obtained as a nearly pure viscous oil. NMR (CDCl₃): δ8.60 (1H,d, J=5 Hz), 7.69-7.72 (2H, m), 7.23-7.35 (6H, m), 7.17 (2H, br d, J=7Hz), 6.98 (1H, s), 6.61 (1H, s),4.22 (3H, s), 3.52 (2H, s), 2.95 (2H, d,J=12 Hz), 2.72 (1H, m), 2.10 (2H, br t, J=11 Hz), 1.97 (2H, br d, J=12Hz), 1.81 (2H, br q, J=11).

This material was re-chromatographed on a HPLC by reversed-phase elutionwith 95 to 5% water/CH₃CN(0.1% TFA) to obtain pure title compound (2-3)as a TFA salt. The pure fractions were combined, the solvents evaporatedand the residue chased with toluene to remove traces of water. Uponstirring this residue in diethyl ether, the product slowly precipitatedas a white solid (60 mg). m/e(m+1): 503.2

A mixture of3-methoxy-5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-pyrazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazine(2-3) TFA (36.8 mg, 0.059 mmol) and pyridine hydrochloride (591 mg, 5.1mmol) was heated at 150° C. for 5 min. This solid was dissolved inwater, the solution made basic with NaHCO₃, and the product extractedinto methylene chloride/methanol (5%). This solution was dried, filteredand the solvents evaporated. This residue was digested in acetonitrileto give5-phenyl-6-(4-{[4-(5-pyridin-2-yl-1H-pyrazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazin-3-ol(2-4) as a tan solid freebase. NMR (CDCl₃/CD3OD): δ8.66 (1H, br d, J=4.5Hz), 8.40 (1H, br d, J=7 Hz), 8.29 (1H, br m), 7.67 (2H, br m), 7.55(2H, d, J=7.5 Hz), 7.42 (1H, s), 7.35 (1H, t, J=7.5 Hz), 7.11-7.30 (4H,m), 7.08 (2H, d, J=9 Hz), 6.98 (1H, s), 4.21 (2H, s), 3.47 (2H, d, J=10Hz), 3.26 (1H, m), 3.00 (2H, m), 2.43 (2H, br d, J=11 Hz), 2.13 (2H, m);m/e (m+1): 489.2.

4-phenyl-6-pyridin-3-yl-3-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazine(3-4)

4-Phenyl-3,6-dichloropyridazine (1-1; 113 mg, 0.5 mmol) was dissolved indry dioxane (2 mL). Then pyridine-3-boronic acid (99 mg, 0.8 mmol), 2 Maqueous Na₂CO₃ (0.75 mL) and palladium(0) tetrakis-triphenylphosphine(60 mg, 0.04 mmol) were added. This mixture was degassed, the vesselsealed and heated at 110° C. for 9 hours. The cooled reaction wasdiluted with water and the product mixture extracted into ethyl acetate,the extract dried over anhydrous NaSO₄, filtered and evaporated. Theproduct was purified by chromatography on silica gel and elution with a20-70% ethyl acetate/hexane gradient to give3-chloro-4-phenyl-6-pyridin-3-ylpyridazine (3-1). NMR (CDCl₃): δ9.26(1H, s), 8.78 (1H, d, J=4.6 Hz), 8.51 (1H, d, J=8 Hz), 7.86 (1H, s),7.55-7.58 (5H, m), 7.51 (1H, dd, J=8 Hz, J=4.5 Hz); m/e (m+1): 268.1.

4-(4-Phenyl-6-pyridin-3-ylpyridazin-3-yl)benzaldehyde (3-2), wasprepared according to the procedure described in Scheme 1, except3-chloro-4-phenyl-6-pyridin-3-ylpyridazine (3-1) is substituted forN,N-dimethyl 6-chloro-5-phenylpyridazin-3-amine (1-2). NMR (CDCl₃):δ10.06 (1H, s), 9.36 (1H,s), 8.78 (1H, d, J=4.6 Hz), 8.60 (1H, d, J=11.7Hz), 7.95 (1H, s), 7.85 (2H, d, J=8 Hz), 7.69 (2H, d, J=8 Hz), 7.52 (1H,m), 7.38-7.44 (3H, m), 7.28 (2H, d, J=4.6 Hz); m/e (m+1): 338.2

4-Phenyl-6-pyridin-3-yl-3-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyridazine(3-4) was prepared according to the procedure described in Scheme 1,except 4-(4-phenyl-6-pyridin-3-ylpyridazin-3-yl)benzaldehyde (3-2) issubstituted for 4-[6-(dimethylamino)-4-phenylpyridazin-3-yl]benzaldehyde(1-3) and 2-(3-piperidin-4-yl-1H-1,2,4-triazol-5-yl)pyridine (3-3; seesynthesis below) for 2-(3-piperidin-4-yl-1H-pyazol-5-yl)pyridine (1-4).NMR (CDCl₃): δ9.34 (1H, s), 8.76 (1H, d, J=4 Hz), 8.64 (1H, d, J=4.3Hz), 8.58 (1H, d, J=8 Hz), 8.14 (1H, d, J=7.8 Hz), 7.88 (1H, s), 7.83(1H, t, J=7.8 Hz), 7.44-7.53 (3H, m), 7.30-7.42 (5H, m), 3.57 (2H, s),2.98 (2H, br s), 2.87 (1H, m), 2.06-2.22 (4H, m), 2.00 (2H, m); m/e(m+1): 551.5

2-(3-Piperidin-4-yl-1H-1,2,4-triazol-5-yl)pyridine (3-3)

Carbonyl diimidazole (3.57 g, 22 mmol) was added to a solution of1-(tert-butoxycarbonyl)-piperidine-4-carboxylic acid (4.59 g, 20 mmol)in methylene chloride (50 mL) and stirred for two hours until gasevolution ceased. Then hydrazine (0.8 mL, ±26 mmol) was added to thereaction and the reaction was stirred at room temperature for anothertwo hours. The reaction was diluted with more methylene chloride andwashed with sat'd aqueous NaHCO₃. The organic layer was dried overanhydrous Na₂SO₄, filtered and the solvent evaporated to give a viscousresidue. Trituration with diethyl ether afforded tert-butyl4-(hydrazinocarbonyl)-piperidine-1-carboxylate as an off-white solid.NMR (CDCl₃): δ6.77 (1H, br s), 4.15 (2H, br s), 3.90 (2H, v br s), 2.75(2H, b s), 2.22 (1H, m), 1.78 (2H, br d, J=11.9 Hz), 1.66 (2H, br q,J=12.2 Hz, J=27.5 Hz), 1.47 (9H, s).

This material (2.43 g, 10 mmol) was dissolved in anhydrous2-ethoxyethanol (20 mL) and 2-cyanopyridine (1.14 g, 11 mmol) was addedto the solution. After 25 wt. % sodium methoxide/methanol (1.1 mL, ˜5mmol) was added the mixture was heated to 130° C. for 16 hours. Thecooled reaction was neutralized with acetic acid and partitioned betweenethyl acetate and aq. NaHCO₃. The organic layer was dried over Na₂SO₄,the salts removed by filtration and the solvent evaporated under vacuum.This residue was triturated with diethyl ether to give tert-butyl4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidine-1-carboxylate as awhite solid. NMR (CDCl₃): δ8.70 (1H, d, J=3.9 Hz), 8.19 (1H, d, J=7.9Hz), 7.87 (1H, d t, J=1.7 Hz, J=8 Hz), 7.40 (1H, m), 4.20 (2H, br s),3.03 (1H, m), 2.95 (2H, br s), 2.09 (2H, br d, J=12 Hz), 1.86 (2H, br q,J=4.2 Hz), 1.49 (9H, s); rn/e (m+1): 330.2

This material (2.68 g, 8.14 mmol) was suspended in 4 N HCl/dioxane. Thestoppered reaction mixture was stirred at room temperature for 16 hoursand then diluted with diethyl ether. The solids were isolated byfiltration and the hydroscopic solid was digested in acetonitrile. Thissolid was isolated by filtration and partially dissolved in hotmethanol. Upon cooling and addition of some ethyl ether to the mixture2-(3-piperidin-4-yl-1H-1,2,4-triazol-5-yl)pyridine (3-3) was obtained asthe dihydrochloride salt. NMR (DMSO-d6): δ9.10 (1H, br s), 8.92 (1H, brs), 8.73 (1H, d, J=4.9 Hz), 8.10-8.20 (2H, m), 7.64 (1H, t, J=5.7 Hz),3.33 (2H, br d, J=12.7 Hz), 3.16 (1H, m), 3.05 (2H, br q, J=11.9 Hz,J=21.8 Hz), 2.18 (2H, br d, J=11.5 Hz), 1.99 (2H, br q, j=11.0 Hz,J=22.2 Hz): m/e (m+1): 230.3.

5-phenyl-2-pyridin-2-yl-4-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidine(4-4)

Methyl (2Z)-3-methoxy-3-phenylacrylate (J. Chem. Soc. Perkin Trans. 2,1988, 227-234, 1.00 g, 5.20 mmol), 2-pyridine-2-carboximidamidehydrochloride (0.983 g, 6.24 mmol) and sodium hydroxide (0.250 g, 6.24mmol) were stirred in 10 ml EtOH and heated to reflux. After 1 h thereaction was cooled and concentrated in vacuo. The resulting residue wasdiluted with water, extracted 3× w/EtOAc. The combined extracts wasdried over Na₂SO₄, filtered and concentrated. Purification by flashcolumn chromatography (suspend material on silica, elute with 98:2DCM/MeOH) afforded an impure yellow solid. The solid was triturated withether, filtered, and washed with ether. This afforded5-phenyl-2-pyridin-2-ylpyrimidin-4-ol (4-1) as a white solid. ¹H NMR(CDCl₃): δ 11.22 (bs, 1H), 8.49 (d, J=4.9 Hz, 1H), 8.45 (d, J=4.8 Hz,1H), 8.24 (s, 1H), 7.93 (dt, J=1.7, 7.6 Hz, 1H), 7.77 (d, J=7.1 Hz, 2H),7.50 (ddd, J=1.1, 4.6, 7.6 Hz, 1H), 7.47 (t, J=7.6 Hz, 2H), 7.40 (t,J=7.6 Hz, 1H).

5-Phenyl-2-pyridin-2-ylpyrimidin-4-ol (4-1; 206 mg, 0.826 mmol) wasstirred in 4 ml of anhydrous MeCN. Phosphorous oxychloride (0.077 ml,0.83 mmol) was added and the resulting mixture was heated to reflux.After 1 h the reaction was cooled and concentrated in vacuo and quenchedwith half-saturated aqueous NaHCO₃. The mixture was extracted 3× withDCM. The combined organic phases was dried over Na₂SO₄, filtered andconcentrated to provide 4-chloro-5-phenyl-2-pyridin-2-ylpyrimidine(4-2). 1H NMR (CDCl₃): δ 8.88 (s, 1H), 8.84 (s, 1H), 8.57 (d, J=8.0 Hz,1H), 7.90 (dt, J=1.5, 7.6 Hz, 1H), 7.53 (m, 5H), 7.46 (dd, J=4.9, 7.1Hz, 1H).

4-Chloro-5-phenyl-2-pyridin-2-ylpyrimidine (4-2; 0.15 g, 0.56 mmol),(4-formylphenyl)boronic (0.126 g, 0.840 mmol),bis(tri-t-butylphosphine)palladium(0) (0.014 g, 0.028 mmol) andanhydrous KF (0.107 g, 1.85 mmol) were stirred in 2 ml anhydrous dioxaneunder N2. The reaction was heated to reflux. After 15 min an additionalsample of bis(tri-t-butylphosphine)palladium(0) (0.014 g, 0.028 mmol)was added and the reaction was heated at reflux overnight. The reactionwas cooled and diluted with water. The mixture was extracted 3× withDCM. The combined organic phases was dried over Na₂SO₄, filtered andconcentrated. The residue was purified by flash column chromatography(dissolved sample in DCM, eluted with 98:2 DCM/EA) and provided animpure sample of 4-(5-phenyl-2-pyridin-2-ylpyrimidin-4-yl)benzaldehyde(4-3) that was used directly in the next transformation.

4-(5-Phenyl-2-pyridin-2-ylpyrimidin-4-yl)benzaldehyde (4-3; 0.011 g,0.033 mmol), sodium triacetoxyborohydride (0.010 g, 0.049 mmol) and2-(3-piperidin-4-yl-1H-1,2,4-triazol-5-yl)pyridine hydrochloride (0.022g, 0.082 mmol) were stirred in 0.5 ml of anhydrous DMF. Triethylamine(0.014 ml, 0.098 mmol) and acetic acid (0.007 ml, 0.13 mmol) were addedand the reaction was stirred at ambient temperature for 6 h. Thereaction was quenched by the addition of 3 drops of water and theresulting solution was purified by reverse phase preparativechromatography. This afforded5-phenyl-2-pyridin-2-yl-4-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidine(4-4) as a trifluoroacetic acid salt. ¹H NMR (CDCl₃): δ 9.12 (s, 1H),9.09 (d, 1H), 8.94 (d, 1H), 8.72 (d, 2H), 8.20 (m, 1H), 8.15 (m, 2H),7.98 (s, 1H), 7.75 (d, 2H), 7.58 (m, 3H), 7.43 (m, 2H), 7.36 (m, 2H),4.40 (s, 2H), 3.62 (m, 2H), 3.25 (m, 2H), 2.42 (m, 2H), 2.12 (m, 2H).m/z (m+1): 551.2

The following compound in Table 2 was made in an analogous manner.

TABLE 2 Cmpd. No. Structure Nomenclature MS M + 1 4-5

5-phenyl-4-(4-{[4-(5- pyridin-2-yl-1H-1,2,4- triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidin- 2-amine 488.2

1-{-4-[5-phenyl-2-(pyridine-3-ylamino)pyrimidin-4-yl]benzyl}-4-(5-pyridin-2-yl-4H-1,2,4-triazol-3-yl)piperidine(5-6) and1-{4-[2-(5-amino-1,3,4-thiadiazol-2-yl)-5-phenylpyrimidin-4-yl]benzyl}-4-(5-pyridin-2-yl-4H-1,2,4-triazol-3-yl)piperidine(5-7)

To a solution of 4-cyanobenzaldehyde (5-1) (20.0 g, 152 5 mmol) andethylene glycol (25.5 mL, 457.5 mmol) in toluene (250 mL) was addedp-toluenesulfonic acid (300 mg). The flask was equipped with aDean-Stark trap and the mixture heated to reflux. After 5 hours, themixture was concentrated. The residue was taken up in ethyl acetate andwashed with saturated NaHCO₃, H₂O (2×), and brine. The organic layer wasdried (Mg50₄), filtered, and concentrated to give4-(1,3-dioxolan-2-yl)benzonitrile as a clear oil which solidified undervacuum: 1H-NMR (500 MHz, CDCl₃) δ 7.67 (d, J=8.06 Hz, 2H), 7.59 (d,J=8.30 Hz, 2H), 5.85 (s, 1H), 4.13-4.05 (m, 4H).

To a solution of 4-(1,3-dioxolan-2-yl)benzonitrile (26.9 g, 154 mmol) inanhydrous THF (300 mL) was added benzylmagnesium chloride (100 mL, 2.0 Msolution in THF, 200 mmol) slowly with internal temperature <−30° C.After 1 hr the mixture was warmed to room temperature. After 4 hours,the mixture was recooled to −40° C. and quenched with saturated ammoniumchloride. The mixture was warmed to room temperature and extracted withethyl acetate (3×). The combined organic layers were dried with sodiumsulfate, filtered, and concentrated. Flash column (10%-40% ethylacetate/hexanes) gave the acetal-ketone (5-2) as a pale yellow solid:1H-NMR (500 MHz, CDCl₃) δ 8.02 (d, J=8.30 Hz, 2H), 7.57 (d, J=8.30 Hz,2H), 7.38-7.24 (m, 5H), 5.86 (s, 1H), 4.29 (s, 2H), 4.28-4.09 (m, 4H).

A flask containing acetal-ketone (5-2) (6.7 g, 25 mol),dimethylformamide-dimethylacetal (10 g) and dimethylformamide (2 g) wasfitted with an air cooled condenser and placed in an oil bath at 100° C.to stir for 4.5 hours at which time LCMS indicated that only a trace ofstarting material remained. The mixture was then cooled to 40° C. andisopropyl alcohol (10 ml) was added and allowed to continue stirring.When the mixture reached room temperature, diethyl ether (30 ml) wasadded and the stirring was continued as the mixture was cooled to 0° C.The yellowish crystals of eneaminone (5-3) were filtered off to give5.37 g of product consisted of a mixture of double bond isomers: LCMS,m/e 324.16 (M+H⁺).

To a mixture of eneaminone (5-3) (3.23 g, 10 mmol), thiourea (912 mg, 12mmol) and ethanol (40 ml) was added potassium tert-butoxide (1400 mg,12.5 mmol). The mixture was heated to reflux with stirring for 1 hour atwhich time LCMS indicated that all the starting material had beenconsumed. The reaction was diluted with dichloromethane (50 ml) andcooled to 0° C. Methyl iodide (0.934 ml, 15 mmol) was added at such arate as to maintain an internal temperature <4° C. The reaction mixturewas allowed to stir for 30 minutes during which time a white precipitateforms. The volatiles were removed with a rotary evaporator and theresidue was taken up in ethyl acetate, washed with water, dried withsodium sulfate and concentrated with a rotary evaporator. The resultingresidue was triturated with dichloromethane, diethyl ether and hexanesto yield 3.1 g of a pale yellow solid. The mother liquors could bepurified by flash chromatography to yield another 257 mg of product:LCMS, m/e 351.06 (MAT). A solution of sulfide (3.034 g, 8 6 mmol) indichloromethane (30 ml) was cooled in an ice bath with stirring.Peracetic acid (2.7 g of 32% in acetic acid diluted with 10 mldichloromethane) was added drop-wise at such a rate as to maintain thetemperature below 5° C. Stirring was continued for 2 hours at which timeLCMS indicated that all the starting material had been consumed. Thereaction was quenched with saturated sodium sulfite, washed withsaturated sodium bicarbonate, dried with sodium sulfate and concentratedwith a rotary evaporator to give the sulfoxide (5-4) as a white solid:¹H NMR (500 MHz, CDCl₃). δ 8.86 (s, 1H), 7.52-7.50 (m, 2H), 7.41-7.38(m, 5H), 7.25-7.23 (m, 2H), 5.79 (s, 1H), 4.11-4.01 (m, 4H), 3.04 (s,3H), LCMS, m/e 367.56 (M+H⁺).

A solution of sulfoxide (5-4) (3.1 g, 8 7 mmol) in ethyl acetate (20 ml)was cooled to 0° C. with rapid stirring. Sodium tungstenate (696 mg, 2.1mmol) and hydrogen peroxide (20 ml, 169 mmol) were added and the mixturewas allowed to slowly warm to room temperature with stirring. Thereaction was stirred overnight at which time LCMS indicated that all thesulfoxide had been oxidized to the sulfone. The solution was carefullyquenched with saturated sodium sulfite, dried with sodium sulfate andconcentrated with a rotary evaporator to give the sulfone (5-5) as awhite solid: 1H-NMR (500 MHz, CDCl₃). δ 8.88 (s, 1H), 7.56-7.54 (m, 2H),7.45-7.42 (m, 5H), 7.27-7.25 (m, 2H), 5.82 (s, 1H), 4.14-4.04 (m, 4H),3.46 (s, 3H), LCMS, m/e 383.44 (M+H⁺).

To a solution of sulfone (5-5) (177 mg, 0.48 mmol) and 2 amino-pyridine(64 mg, 0.68 mmol) in THF (4.5 ml) was added sodiumbis(trimethylsilyl)amide (0.360 ml, 0.72 mmol, 2 M in THF). LCMSindicates reaction is done after a few minutes. LCMS m/e 397.69 (M+H⁺).Concentrated HCl (0.5 ml) was then added to the reaction mixture andstirred at room temperature for a few minutes. LCMS indicated that thereaction was complete. LCMS m/e 353.51 (M+H⁺). The volatiles wereremoved on a rotary evaporator, made alkaline with 50% sodium hydroxideand extracted with ethyl acetate (X2). Separately, a solution of thetriazole-amine trifluoroacetate salt in dichloromethane (2 ml) andmethanol (2 ml) was neutralized with Hunig's base (2 ml) and then addedto the organic fraction. The mixture was concentrated using a rotaryevaporator and diluted with dichloromethane (10 ml). The mixture wasstirred for 15 minutes at room temperature and then sodiumtriacetoxyborohydride (405 mg, 1.92 mol) was added. Stirring wascontinued for an additional 18 hours. The reaction mixture was dilutedwith sodium hydroxide (50%, 10 ml) and extracted twice with ethylacetate, dried with sodium sulfate and concentrated on a rotaryevaporator. The residue was purified by preparative reverse phase HPLCto give the product (5-6) as an amorphous solid: ¹H-NMR 0.3 (500 MHz,CD₃OD) 8.80 (s, 1H), 8.75 (br, 1H), 8.43-8.32 (m, 4H), 7.82 (m, 1H)7.65-7.58 (m, 3H), 7.54-7.52 (m, 2H), 7.45-7.42 (m, 1H), 7.40-7.38 (m,3H), 7.28 (m, 2H), 4.40 (s, 2H), 3.64-3.45 (m, 3H), 3.24-3.22 (m, 2H),2.41-2.38 (m, 2H), 2.17-2.15 (m, 2H), LCMS m/e 566.70 (M+H⁺).

A solution of sulfone (5-5) (250 mg, 0.654 mmol) and potassium cyanide(64 mg, 0.983 mmol) in DMSO (5 mL) was stirred at rt for 1 h. Thereaction was diluted with ethyl acetate (50 mL) and washed with water(3×25 mL), and brine (25 mL). The organic layer was dried over magnesiumsulfate and concentrated under reduced pressure. The crude material waspurified by column chromatography (0-35% ethyl acetate/hexanes) to yield4-[4-(1,3-dioxolan-2-yl)phenyl]-5-phenylpyrimidine-2-carbonitrile as awhite solid. ¹H NMR (CDCl₃). δ 8.78 (s, 1H), 7.50 (m, 2H), 7.40 (m, 5H),7.22 (m, 2H), 5.79 (s, 1H), 4.07 (m, 4H); MS (EI) m/z 330 (M+1).

A solution of4-[4-(1,3-dioxolan-2-yl)phenyl]-5-phenylpyrimidine-2-carbonitrile (165mg, 0.501 mmol) in THF (7 mL) and 6N HCl (4 mL) was stirred at rt for 1h. The reaction mixture was neutralized with sodium hydroxide anddiluted with ethyl acetate (50 mL). The solution was washed with water(2×25 mL) and dried over magnesium sulfate. Hunig's base (1 mL) wasadded to the solution and it was then added to a solution of4-(5-pyridin-2-yl-4H-1,2,4-triazol-3-yl)piperidinium trifluoroacetate(206 mg, 0.600 mmol) in dichloromethane (5 mL) and Hunig's base (1 mL).The reaction mixture was concentrated under reduced pressure and dilutedwith dichloromethane (10 mL). Sodium triacetoxyborohydride (320 mg,1.509 mmol) was added and stirred over night. The reaction mixture wasbasified with sodium hydroxide, extracted with ethyl acetate (50 mL)_(j)and purified on reverse phase HPLC to yield5-phenyl-4-(4-{[4-(5-pyridin-2-yl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidine-2-carbonitrileas a solid. MS (EI) m/z 499 (M+1).

A solution of5-phenyl-4-(4-{[4-(5-pyridin-2-yl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidine-2-carbonitrile(100 mg, 0.163 mmol) and hydrazinecarbothioamide (23 mg, 0.252 mmol) intrifluoroacetic acid (2.5 mL) was heated at 75° C. for 15 min.Trifluoroacetic acid was removed under reduced pressure and the crudematerial was purified on reverse phase HPLC to yield the desired product(5-7) (75 mg, 67%). ¹H-NMR (500 MHz, CD₃OD)

8.88 (s, 1H), 8.75 (br s, 1H), 8.35 (m, 2H), 7.28-7.80 (m, 10H), 4.40(s, 2H), 3.23-3.65 (m, 3H), 2.41-2.17 (m, 4H); MS (EI) m/z 573 (M+1).

The following compounds in Table 3 were made in an analogous manner.

TABLE 3

Cmpd. No. R Nomenclature MS M + 1 5-8

1-methyl-4-(2-{[5-phenyl-4-(4-{[4-(5-pyridin-2-yl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidin-2- yl]amino}ethyl)piperazine 614.8 5-9

1-[4-(2-amino-5-phenylpyrimidin-4-yl)benzyl]-4-(5-pyridin-2-yl-4h-1,2,4-triazol- 3-yl)piperidine 488.65-10

1-{4-[2-(methylthio)-5-phenylpyrimidin-4-yl]benzyl}-4-(5-pyridin-2-yl-4h-1,2,4-triazol- 3-yl)piperidine 519.75-11

1-{4-[2-(4-acetylpiperazin-1-yl)-5-phenylpyrimidin-4-yl]benzyl}-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine 599.7 5-12

1-[4-(2-{[1-(ethoxycarbonyl)piperidin-4-yl]amino}-5-phenylpyrimidin-4-yl)benzyl]-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3- yl)piperidine 643.8 5-13

1-methyl-4-[5-phenyl-4-(4-{[4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidinium-1-yl]methyl}phenyl)pyrimidin-2-yl]piperazine 571.7 5-14

1-(2-hydroxyethyl)-4-[5-phenyl-4-(4-{[4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidin-2- yl]piperazine 601.8 5-15

1-[2-(dimethylamino)ethyl]-4-[5-phenyl-4-(4-{[4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidin- 2-yl]piperazine 628.8 5-16

1-(4-{5-phenyl-2-[(pyridin-3- ylmethyl)amino]pyrimidin-4-yl}benzyl)-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3- yl)piperidine 579.7 5-17

2-{[5-phenyl-4-(4-{[4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidin-1- yl]methyl}phenyl)pyrimidin-2-yl]amino}pyridine 565.7 5-18

2-{5-[1-(4-{2-[methyl(pyridin-2-yl)amino]-5-phenylpyrimidin-4-yl}benzyl)piperidin-4-yl]-4h-1,2,4-triazol-3-yl}pyridine 579.7 5-19

1-{4-[2-(dimethylamino)-5-phenylpyrimidin-4-yl]benzyl}-4-(5-pyridin-2-yl-4h-1,2,4- triazol-3-yl)piperidine 516.75-20

1-(4-{2-[[2-(dimethylamino)ethyl](methyl)amino]-5-phenylpyrimidin-4-yl}benzyl)-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3- yl)piperidine 573.8 5-21

1-[4-(2-{[3-(dimethylammonio) propyl]amino}-5-phenylpyrimidin-4-yl)benzyl]-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3- yl)piperidine 573.8 5-22

1-[4-(2-{[2-(dimethylammonio)ethyl]amino}-5-phenylpyrimidin-4-yl)benzyl]-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3- yl)piperidine 559.7 5-23

1-(4-{2-[[3-(dimethylammonio) propyl](methyl)amino]-5-phenylpyrimidin-4-yl}benzyl)-4-(5-pyridin-2-yl-4h-1,2,4- triazol-3-yl)piperidine 587.8

Example 1 Cloning of the Human Akt Isoforms and APH-Akt1

The pS2neo vector (deposited in the ATCC on Apr. 3, 2001 as ATCCPTA-3253) was prepared as follows: The pRmHA3 vector (prepared asdescribed in Nucl. Acid Res. 16:1043-1061 (1988)) was cut with BglII anda 2734 by fragment was isolated. The pUChsneo vector (prepared asdescribed in EMBO J. 4:167-171 (1985)) was also cut with BglII and a4029 by band was isolated. These two isolated fragments were ligatedtogether to generate a vector termed pS2neo-1. This plasmid contains apolylinker between a metallothionine promoter and an alcoholdehydrogenase poly A addition site. It also has a neo resistance genedriven by a heat shock promoter. The pS2neo-1 vector was cut with PspSIIand BsiWI. Two complementary oligonucleotides were synthesized and thenannealed (CTGCGGCCGC (SEQ.ID.NO.: 1) and GTACGCGGCCGCAG (SEQ.ID.NO.:2)). The cut pS2neo-1 and the annealed oligonucleotides were ligatedtogether to generate a second vector, pS2neo. Added in this conversionwas a NotI site to aid in the linearization prior to transfection intoS2 cells.

Human Akt1 gene was amplified by PCR (Clontech) out of a human spleencDNA (Clontech) using the 5′ primer:5′CGCGAATTCAGATCTACCATGAGCGACGTGGCTATTGTG 3′ (SEQ.ID.NO.: 3), and the 3′primer: 5′CGCTCTAGAGGATCCTCAGGCCGTGCTGCTGGC3′ (SEQ.ID.NO.: 4). The 5′primer included an EcoRI and BglII site. The 3′ primer included an XbaIand BamHI site for cloning purposes. The resultant PCR product wassubcloned into pGEM3Z (Promega) as an EcoRI/Xba I fragment. Forexpression/purification purposes, a middle T tag was added to the 5′ endof the full length Akt1 gene using the PCR primer:5′GTACGATGCTGAACGATATCTTCG 3′ (SEQ.ID.NO.: 5). The resulting PCR productencompassed a 5′ KpnI site and a 3′ BamHI site which were used tosubclone the fragment in frame with a biotin tag containing insect cellexpression vector, pS2neo.

For the expression of a pleckstrin homology domain (PH) deleted (Δaa4-129, which includes deletion of a portion of the Akt1 hinge region)version of Akt1, PCR deletion mutagenesis was done using the full lengthAkt1 gene in the pS2neo vector as template. The PCR was carried out in 2steps using overlapping internal primers(5′GAATACATGCCGATGGAAAGCGACGGGGCTGAAGAGATGGAGGTG 3′ (SEQ.ID.NO.: 6), and5′CCCCTCCATCTCTTCAGCCCCGTCGCTTTCCATCGGCATG TATTC 3′ (SEQ.ID.NO.: 7))which encompassed the deletion and 5′ and 3′ flanking primers whichencompassed the KpnI site and middle T tag on the 5′ end. The final PCRproduct was digested with KpnI and SmaI and ligated into the pS2neo fulllength Akt1 KpnI/Sma I cut vector, effectively replacing the 5′ end ofthe clone with the deleted version.

Human Akt3 gene was amplified by PCR of adult brain cDNA (Clontech)using the amino terminal oligo primer: 5′GAATTCAGATCTACCATGAGCGATGTTACCATTGTG 3′ (SEQ.ID.NO.: 8); and the carboxyterminal oligo primer:

(SEQ. ID. NO.: 9) 5′ TCTAGATCTTATTCTCGTCCACTTGCAGAG 3′.These primers included a 5′ EcoRI/BglII site and a 3′ XbaI/BglII sitefor cloning purposes. The resultant PCR product was cloned into theEcoRI and XbaI sites of pGEM4Z (Promega). For expression/purificationpurposes, a middle T tag was added to the 5′ end of the full length Akt3clone using the PCR primer: 5′GGTACCATGGAATACATGCCGATGGAAAGCGATGTTACCATTGTGAAG 3′(SEQ.ID.NO.: 10). Theresultant PCR product encompassed a 5′ KpnI site which allowed in framecloning with the biotin tag containing insect cell expression vector,pS2neo.

Human Akt2 gene was amplified by PCR from human thymus cDNA (Clontech)using the amino terminal oligo primer: 5′AAGCTTAGATCTACCATGAATGAGGTGTCTGTC 3′ (SEQ.ID.NO.: 11); and the carboxyterminal oligo primer: 5′ GAATTCGGATCCTCACTCGCGGATGCTGGC 3′ (SEQ.ID.NO.:12). These primers included a 5′ HindIII/BglII site and a 3′ EcoRI/BamHIsite for cloning purposes. The resultant PCR product was subcloned intothe HindIII/EcoRI sites of pGem3Z (Promega). For expression/purificationpurposes, a middle T tag was added to the 5′ end of the full length Akt2using the PCR primer: 5′GGTACCATGGAATACATGCCGATGGAAAATGAGGTGTCTGTCATCAAAG 3′ (SEQ.ID.NO.: 13).The resultant PCR product was subcloned into the pS2neo vector asdescribed above.

Example 2 Expression of Human Akt Isoforms and APH-Akt1

The DNA containing the cloned Akt1, Akt2, Akt3 and APH-Akt1 genes in thepS2neo expression vector was purified and used to transfect DrosophilaS2 cells (ATCC) by the calcium phosphate method. Pools of antibiotic(G418, 500 μg/ml) resistant cells were selected. Cells were expanded toa 1.0 L volume (˜7.0×10⁶/ml), biotin and CuSO₄ were added to a finalconcentration of 50 μM and 50 mM respectively. Cells were grown for 72 hat 27° C. and harvested by centrifugation. The cell paste was frozen at−70° C. until needed.

Example 3 Purification of Human Akt Isoforms and ΔPH-Akt1

Cell paste from one liter of S2 cells, described in Example 2, was lysedby sonication with 50 mls 1% CHAPS in buffer A: (50 mM Tris pH 7.4, 1 mMEDTA, 1 mM EGTA, 0.2 mM AEBSF, 10 μg/ml benzamidine, 5 μg/ml ofleupeptin, aprotinin and pepstatin each, 10% glycerol and 1 mM DTT). Thesoluble fraction was purified on a Protein G Sepharose fast flow(Pharmacia) column loaded with 9 mg/ml anti-middle T monoclonal antibodyand eluted with 75 μl\A EYMPME (SEQ.ID.NO.: 14) peptide in buffer Acontaining 25% glycerol. Akt containing fractions were pooled and theprotein purity evaluated by SDS-PAGE. The purified protein wasquantitated using a standard Bradford protocol. Purified protein wasflash frozen on liquid nitrogen and stored at −70° C.

Akt and Akt pleckstrin homology domain deletions purified from S2 cellsrequired activation. Akt and Akt pleckstrin homology domain deletionswas activated (Alessi et al. Current Biology 7:261-269) in a reactioncontaining 10 nM PDK1 (Upstate Biotechnology, Inc.), lipid vesicles (10μM phosphatidylinositol-3,4,5-trisphosphate—Metreya, Inc, 100 μMphosphatidylcholine and 100 μM phosphatidylserine—Avanti Polar lipids,Inc.) and activation buffer (50 mM Tris pH7.4, 1.0 mM DTT, 0.1 mM EGTA,1.0 μM Microcystin-LR, 0.1 mM ATP, 10 mM MgCl₂, 333 μg/ml BSA and 0.1 mMEDTA). The reaction was incubated at 22° C. for 4 hours. Aliquots wereflash frozen in liquid nitrogen.

Example 4 Akt Kinase Assays

Activated Akt isoforms and pleckstrin homology domain deletionconstructs were assayed utilizing a GSK-derived biotinylated peptidesubstrate. The extent of peptide phosphorylation was determined byHomogeneous Time Resolved Fluorescence (HTRF) using a lanthanidechelate(Lance)-coupled monoclonal antibody specific for thephosphopeptide in combination with a streptavidin-linked allophycocyanin(SA-APC) fluorophore which will bind to the biotin moiety on thepeptide. When the Lance and APC are in proximity (i.e. bound to the samephosphopeptide molecule), a non-radiative energy transfer takes placefrom the Lance to the APC, followed by emission of light from APC at 665nm.

Materials required for the assay:A. Activated Akt isozyme or pleckstrin homology domain deletedconstruct.B. Akt peptide substrate: GSK3α (S21) Peptide no. 3928biotin-GGRARTSSFAEPG (SEQ.ID.NO.:15), Macromolecular Resources.C. Lance labeled anti-phospho GSK3a monoclonal antibody (Cell SignalingTechnology, clone # 27).D. SA-APC (Prozyme catalog no. PJ25S lot no. 896067).

E. Microfluor®B U Bottom Microtiter Plates (Dynex Technologies, Catalogno. 7205). F. Discovery® HTRF Microplate Analyzer, Packard InstrumentCompany.

G. 100× Protease Inhibitor Cocktail (PIC): 1 mg/ml benzamidine, 0.5mg/ml pepstatin, 0.5 mg/ml leupeptin, 0.5 mg/ml aprotinin.H. 10× Assay Buffer: 500 mM HEPES, pH 7.5, 1% PEG, mM EDTA, 1 mM EGTA,1% BSA, 20 mM ∂-Glycerol phosphate.I. Quench Buffer: 50 mM HEPES pH 7.3, 16.6 mM EDTA, 0.1% BSA, 0.1%Triton X-100, 0.17 nM Lance labeled monoclonal antibody clone # 27,0.0067 mg/ml SA-APCJ. ATP/MgCl₂ working solution: 1× Assay buffer, 1 mM DTT, 1×PIC, 125 mMKCl, 5% Glycerol, 25 mM MgCl₂, 375 ™ ATPK. Enzyme working solution: 1× Assay buffer, 1 mM DTT, 1×PIC, 5%Glycerol, active Akt. The final enzyme concentrations were selected sothat the assay was in a linear response range.L. Peptide working solution: 1× Assay buffer, 1 mM DTT, 1×PIC, 5%Glycerol, 2 ™ GSK3 biotinylated peptide # 3928

The reaction is assembled by adding 16 TL of the ATP/MgCl₂ workingsolution to the appropriate wells of a 96-well microtiter plate.Inhibitor or vehicle (1.0 Tl) is added followed by 10 Tl of peptideworking solution. The reaction is started by adding 13 Tl of the enzymeworking solution and mixing. The reaction is allowed to proceed for 50min and then stopped by the addition of 60 Tl HTRF quench buffer. Thestopped reactions were incubated at room temperature for at least 30 minand then read on the Discovery instrument.

PKA Assay:

Each individual PKA assay consists of the following components:

A. 5×PKA assay buffer (200 mM Tris pH7.5, 100 mM MgCl₂, 5 mMO-mercaptoethanol, 0.5 mM EDTA)B. 50 μM stock of Kemptide (Sigma) diluted in waterC. ³³P-ATP prepared by diluting 1.0 μl ³³P-ATP [10 mCi/ml] into 200 Tlof a 50 μM stock of unlabeled ATPD. 10 pa of a 70 nM stock of PKA catalytic subunit (UBI catalog #14-114)diluted in 0.5 mg/ml BSAE. PKA/Kemptide working solution: equal volumes of 5×PKA assay buffer,Kemptide solution and PKA catalytic subunit.

The reaction is assembled in a 96 deep-well assay plate. The inhibitoror vehicle (10 Tl) is added to 10 Tl of the ³³P-ATP solution. Thereaction is initiated by adding 30 Tl of the PKA/Kemptide workingsolution to each well. The reactions were mixed and incubated at roomtemperature for 20 min. The reactions were stopped by adding 50 Tl of100 mM EDTA and 100 mM sodium pyrophosphate and mixing.

The enzyme reaction product (phosphorylated Kemptide) was collected onp81 phosphocellulose 96 well filter plates (Millipore). To prepare theplate, each well of a p81 filter plate was filled with 75 mM phosphoricacid. The wells were emptied through the filter by applying a vacuum tothe bottom of the plate. Phosphoric acid (75 mM, 170 μl) was added toeach well. A 30 μl aliquot from each stopped PKA reaction was added tocorresponding wells on the filter plate containing the phosphoric acid.The peptide was trapped on the filter following the application of avacuum and the filters were washed 5 times with 75 mM phosphoric acid.After the final wash, the filters were allowed to air dry. Scintillationfluid (30 μl) was added to each well and the filters counted on aTopCount (Packard).

PKC Assay:

Each PKC assay consists of the following components:A. 10×PKC co-activation buffer: 2.5 mM EGTA, 4 mM CaCl₂B. 5×PKC activation buffer: 1.6 mg/ml phosphatidylserine, 0.16 mg/mldiacylglycerol, 100 mM Tris pH 7.5, 50 mM MgCl₂, 5 mM ∂-mercaptoethanolC. ³³P-ATP prepared by diluting 1.0 μl ³³P-ATP [10 mCi/ml] into 100 μlof a 100 μM stock of unlabeled ATPD. Myelin basic protein (350 μg/ml, UBI) diluted in waterE. PKC (50 ng/ml, UBI catalog # 14-115) diluted into 0.5 mg/ml BSAF. PKC/Myelin Basic Protein working solution: Prepared by mixing 5volumes each of PKC co-activation buffer and Myelin Basic protein with10 volumes each of PKC activation buffer and PKC.

The assays were assembled in 96 deep-well assay plates Inhibitor orvehicle (10 Tl) was added to 5.0 ul of ³³P-ATP. Reactions were initiatedwith the addition of the PKC/Myelin Basic Protein working solution andmixing. Reactions were incubated at 30° C. for 20 min. The reactionswere stopped by adding 50 Tl of 100 mM EDTA and 100 mM sodiumpyrophosphate and mixing. Phosphorylated Mylein Basic Protein wascollected on PVDF membranes in 96 well filter plates and quantitated byscintillation counting.

Specific compounds of the instant invention were tested in the assaydescribed above and were found to have IC₅₀ of ≦50 μM against one ormore of Akt1, Akt2 and Akt3.

Example 5 Cell based Assays to Determine Inhibition of Akt

Cells (for example LnCaP or a PTEN^((−/−)) tumor cell line withactivated Akt) were plated in 100 mM dishes. When the cells wereapproximately 70 to 80% confluent, the cells were refed with 5 mls offresh media and the test compound added in solution. Controls includeduntreated cells, vehicle treated cells and cells treated with eitherLY294002 (Sigma) or wortmanin (Sigma) at 20 μM or 200 nM, respectively.The cells were incubated for 2, 4 or 6 hrs and the media removed. Thecells were washed with PBS, scraped and transferred to a centrifugetube. They were pelleted and washed again with PBS. Finally, the cellpellet was resuspended in lysis buffer (20 mM Tris pH8, 140 mM NaCl, 2mM EDTA, 1% Triton, 1 mM Na Pyrophosphate, 10 mM 8-Glycerol Phosphate,10 mM NaF, 0 5 mm NaVO₄, 1 μM Microsystine, and 1× Protease InhibitorCocktail), placed on ice for 15 minutes and gently vortexed to lyse thecells. The lysate was spun in a Beckman tabletop ultra centrifuge at100,000×g at 4° C. for 20 min. The supernatant protein was quantitatedby a standard Bradford protocol (BioRad) and stored at 70° C. untilneeded.

Proteins were immunoprecipitated (IP) from cleared lysates as follows:For Akt1, lysates are mixed with Santa Cruz sc-7126 (D-17) in NETN (100mM NaCl, 20 mM Tris pH 8.0, 1 mM EDTA, 0.5% NP-40) and Protein A/GAgarose (Santa Cruz sc-2003) was added. For Akt2, lysates were mixed inNETN with anti-Akt-2 agarose (Upstate Biotechnology #16-174) and forAkt3, lysates were mixed in NETN with anti-Akt3 agarose (UpstateBiotechnology #16-175). The IPs were incubated overnight at 4° C.,washed and seperated by SDS-PAGE.

Western blots were used to analyze total Akt, pThr308 Akt1, pSer473Akt1, and corresponding phosphorylation sites on Akt2 and Akt3, anddownstream targets of Akt using specific antibodies (Cell SignalingTechnology): Anti-Total Akt (cat. no. 9272), Anti-Phopho Akt Serine 473(cat. no. 9271), and Anti-Phospho Akt Threonine 308 (cat. no. 9275).After incubating with the appropriate primary antibody diluted inPBS+0.5% non-fat dry milk (NFDM) at 4° C. overnight, blots were washed,incubated with Horseradish peroxidase (HRP)-tagged secondary antibody inPBS+0.5% NFDM for 1 hour at room temperature. Proteins were detectedwith ECL Reagents (Amersham/Pharmacia Biotech RPN2134).

Example 6 Heregulin Stimulated Akt Activation

MCF7 cells (a human breast cancer line that is PTEN^(+/+)) were platedat 1×10⁶ cells per 100 mM plate. When the cells were 70-80% confluent,they were refed with 5 ml of serum free media and incubated overnight.The following morning, compound was added and the cells were incubatedfor 1-2 hrs, after which time heregulin was added (to induce theactivation of Akt) for 30 minutes and the cells were analyzed asdescribed above.

Example 7 Inhibition Of Tumor Growth

In vivo efficacy of an inhibitor of the growth of cancer cells may beconfirmed by several protocols well known in the art.

Human tumor cell lines which exhibit a deregulation of the PI3K pathway(such as LnCaP, PC3, C33a, OVCAR-3, MDA-MB-468 or the like) are injectedsubcutaneously into the left flank of 6-10 week old female nude mice(Harlan) on day 0. The mice are randomly assigned to a vehicle, compoundor combination treatment group. Daily subcutaneous administration beginson day 1 and continues for the duration of the experiment.Alternatively, the inhibitor test compound may be administered by acontinuous infusion pump. Compound, compound combination or vehicle isdelivered in a total volume of 0.2 ml. Tumors are excised and weighedwhen all of the vehicle-treated animals exhibited lesions of 0.5-1.0 cmin diameter, typically 4 to 5.5 weeks after the cells were injected. Theaverage weight of the tumors in each treatment group for each cell lineis calculated.

1. A compound of the Formula A-3:

wherein: X is CH and Y is N; Q is: R¹; a is 0 or 1; b is 0 or 1; m is 0,1, or 2; n is 0, 1, 2, 3 or 4; p is 0, 1, 2, 3,4 or 5; R¹ isindependently selected from: H, (C═O)_(a)O_(b)C₁-C₁₀ alkyl,(C═O)_(a)O_(b) aryl, (C═O)_(a)O_(b)C₂-C₁₀ alkenyl, (C═O)_(a)O_(b)C₂-C₁₀alkynyl, CO₂H, halo, OH, O_(b)C₁-C₆ perfluoroalkyl, (C═O)_(a)NR⁷R⁸, CN,(C═O)_(a)O_(b)C₃-C₈ cycloalkyl, S(O)_(m)NR⁷R⁸, S(O)_(m)-(C₁-C₁₀)alkyland (C═O)_(a)O_(b)heterocyclyl, said alkyl, aryl, alkenyl, alkynyl,cycloalkyl, and heterocyclyl is optionally substituted with one or moresubstituents selected from R⁶; R²′ is independently selected from: H,(C═O)_(a)O_(b)C₁-C₁₀ alkyl, (C═O)_(a)O_(b) aryl, (C═O)_(a)O_(b)C₂-C₁₀alkenyl, (C═O)_(a)O_(b)C₂-C₁₀ alkynyl, CO₂H, halo, OH, O_(b)C₁-C₆perfluoroalkyl, (C═O)_(a)NR⁷R⁸, CN, (C═O)_(a)O_(b)C₃-C₈ cycloalkyl,S(O)_(m)NR⁷R⁸, S(O)_(m)—(C₁-C₁₀)alkyl and (C═O)_(a)O_(b)heterocyclyl,said alkyl, aryl, alkenyl, alkynyl, cycloalkyl, and heterocyclyl isoptionally substituted with one or more substituents selected from R⁶;R⁶ is independently: (C═O)_(a)O_(b)C₁-C₁₀ alkyl, (C═O)_(a)O_(b)aryl,C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, (C═O)_(a)O_(b) heterocyclyl, CO₂H, halo,CN, OH, O_(b)C₁-C₆ perfluoroalkyl, O_(a)(C═O)_(b)NR⁷R⁸, oxo, CHO,(N═O)R⁷R⁸, S(O)_(m)NR⁷R⁸, S(O)_(m)—(C₁-C₁₀)alkyl or (C═O)_(a)O_(b)C₃-C₈cycloalkyl, said alkyl, aryl, alkenyl, alkynyl, heterocyclyl, andcycloalkyl optionally substituted with one or more substituents selectedfrom R^(6a); R^(6a) is independently selected from:(C═O)_(a)O_(b)(C₁-C₁₀)alkyl, O_(a)(C₁-C₃)perfluoroalkyl,(C₀-C₆)alkylene-S(O)_(m)R^(a), oxo, OH, halo, CN, (C₂-C₁₀)alkenyl,(C₂-C₁₀)alkynyl, (C₃-C₆)cycloalkyl, (C₀-C₆)alkylene-aryl,(C₀-C₆)alkylene-heterocyclyl, (C₀-C₆)alkylene-N(R^(b))₂, C(O)R^(a),(C₀-C₆)alkylene-CO₂R^(a), C(O)H, and (C₀-C₆)alkylene-CO₂H, said alkyl,alkenyl, alkynyl, cycloalkyl, aryl, and heterocyclyl is optionallysubstituted with up to three substituents selected from R^(b), OH,(C₁-C₆)alkoxy, halogen, CO₂H, CN, O(C═O)C₁-C₆ alkyl, oxo, and N(R^(b))₂;R⁷ and R⁸ are independently selected from: H, (C═O)O_(b)C₁-C₁₀ alkyl,(C═O)O_(b)C₃-C₈ cycloalkyl, (C═O)O_(b)aryl, (C═O)O_(b)heterocyclyl,C₁-C₆alkyl, aryl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, heterocyclyl, C₃-C₉cycloalkyl, SO₂R^(a), and (C═O)NR^(b) ₂, said alkyl, cycloalkyl, aryl,heterocylyl, alkenyl, and alkynyl is optionally substituted with one ormore substituents selected from R^(6a), or R⁷ and R⁸ can be takentogether with the nitrogen to which they are attached to form amonocyclic or bicyclic heterocycle with 5-7 members in each ring andoptionally containing, in addition to the nitrogen, one or twoadditional heteroatoms selected from N, O and S, said monocyclic orbicyclic heterocycle optionally substituted with one or moresubstituents selected from R^(6a); R^(a) is independently: (C₁-C₆)alkyl,(C₃-C₆)cycloalkyl, aryl, or heterocyclyl; and R^(b) is independently: H,(C₁-C₆)alkyl, aryl, heterocyclyl, (C₃-C₆)cycloalkyl, (C═O)OC₁-C₆ alkyl,(C═O)C₁-C₆ alkyl or S(O)₂R^(a); or a pharmaceutically acceptable salt ora stereoisomer thereof.
 2. The compound according to claim 1 of theFormula A-3: wherein: Q is: (C₃-C₈)cycloalkyl, aryl or heterocyclyl,which is optionally substituted with 1-3 R⁶; and all other substituentsand variables are as defined in claim 1; or a pharmaceuticallyacceptable salt or a stereoisomer thereof.
 3. The compound according toclaim 2 of the Formula B-3: wherein:

Q is selected from:

which are optionally substituted with 1-3 R⁶; n is 0, 1 or 2; and allsubstituents and variables are as defined in claim 2; or apharmaceutically acceptable salt or a stereoisomer thereof.
 4. Thecompound according to claim 3 of the Formula B-3: wherein: Q is selectedfrom:

which are optionally substituted with 1-3 R⁶; and all substituents andvariables are as defined in claim 3; or a pharmaceutically acceptablesalt or a stereoisomer thereof.
 5. A compound which is selected from:5-phenyl-2-pyridin-2-yl-4-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidine;5-phenyl-4-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidin-2-amine;1-{4-[5-phenyl-2-(pyridine-3-ylamino)pyrimidin-4-yl]benzyl}-4-(5-pyridin-2-yl-4H-1,2,4-triazol-3-yl)piperidine;1-{4-[2-(5-amino-1,3,4-thiadiazol-2-yl)-5-phenylpyrimidin-4-yl]benzyl}-4-(5-pyridin-2-yl-4H-1,2,4-triazol-3-yl)piperidine;1-methyl-4-(2-{[5-phenyl-4-(4-{[4-(5-pyridin-2-yl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidin-2-yl]amino}ethyl)piperzine;1-[4-(2-amino-5-phenylpyrimidin-4-yl)benzyl]-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine;1-{4-[2-(methylthio)-5-phenylpyrimidin-4-yl]benzyl}-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine;1-{4-[2-(4-acetylppiperazin-1-yl)-5-phenylpyrimidin-4-yl]benzyl}-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine;1-[4-(2-{[1-(ethoxycarbonyl)piperidin-4-yl]amino}-5-phenylpyrimidin-4-yl)benzyl]-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine;1-methyl-4-[5-phenyl-4-(4-{[4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidin-2-yl]piperazine;1-(2-hydroxyethyl)-4-[5-phenyl-4-(4-{[4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidin-2-yl]piperazine;1-[2-(dimethylamino)ethyl]-4-[5-phenyl-4-(4-{[4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidin-2-yl]piperazine;1-(4-{5-phenyl-2-[(pyridin-3-ylmethyl)amino]pyrimidin-4-yl}benzyl)-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine;2-{[5-phenyl-4-(4-{[4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidin-2-yl]amino}pyridine;2-{5-[1-(4-{2-[methyl(pyridin-2-yl)amino]-5-phenylpyrimidin-4-yl}benzyl)piperidin-4-yl]-4h-1,2,4-triazol-3-yl}pyridine;1-{4-[2-(dimethylamino)-5-phenylpyrimidin-4-yl]benzyl}-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine;1-(4-{2-[[2-(dimethylamino)ethyl](methyl)amino]-5-phenylpyrimidin-4-yl}benzyl)-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine;1-[4-(2-{[3-(dimethylammonio)propyl]amino}-5-phenylpyrimidin-4-yl)benzyl]-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine;1-[4-(2-{[2-(dimethylammonio)ethyl]amino}-5-phenylpyrimidin-4-yl)benzyl]-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine;and1-(4-{2-[[3-(dimethylammonio)propyl](methyl)amino]-5-phenylpyrimidin-4-yl}benzyl)-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine;or a pharmaceutically acceptable salt or a stereoisomer thereof.
 6. TheTFA salt of a compound according to claim 1 which is selected from:5-phenyl-2-pyridin-2-yl-4-(4-{[4-(5-pyridin-2-yl-1H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidine;1-{4-[5-phenyl-2-(pyridine-3-ylamino)pyrimidin-4-yl]benzyl}-4-(5-pyridin-2-yl-4H-1,2,4-triazol-3-yl)piperidine;1-{4-[2-(5-amino-1,3,4-thiadiazol-2-yl)-5-phenylpyrimidin-4-yl]benzyl}-4-(5-pyridin-2-yl-4H-1,2,4-triazol-3-yl)piperidine;1-methyl-4-(2-{[5-phenyl-4-(4-{[4-(5-pyridin-2-yl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidin-2-yl]amino}ethyl)piperzine;1-[4-(2-amino-5-phenylpyrimidin-4-yl)benzyl]-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine;1-{4-[2-(methylthio)-5-phenylpyrimidin-4-yl]benzyl}-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine;1-{4-[2-(4-acetylpiperazin-1-yl)-5-phenylpyrimidin-4-yl]benzyl}-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine;1-[4-(2-{[1-(ethoxycarbonyl)piperidin-4-yl]amino}-5-phenylpyrimidin-4-yl)benzyl]-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine;1-methyl-4-[5-phenyl-4-(4-{[4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidin-2-yl]piperazine;1-(2-hydroxyethyl)-4-[5-phenyl-4-(4-{[4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidin-2-yl]piperazine;1-[2-(dimethylamino)ethyl]-4-[5-phenyl-4-(4-{[4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidin-2-yl]piperazine;1-(4-{5-phenyl-2-[(pyridin-3-ylmethyl)amino]pyrimidin-4-yl}benzyl)-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine;2-{[5-phenyl-4-(4-{[4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)pyrimidin-2-yl]amino}pyridine;2-{5-[1-(4-{2-[methyl(pyridin-2-yl)amino]-5-phenylpyrimidin-4-yl}benzyl)piperidin-4-yl]-4h-1,2,4-triazol-3-yl}pyridine;1-{4-[2-(dimethylamino)-5-phenylpyrimidin-4-yl]benzyl}-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine;1-(4-{2-[[2-(dimethylamino)ethyl](methyl)amino]-5-phenylpyrimidin-4-yl}benzyl)-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine;1-[4-(2-{[3-(dimethylammonio)propyl]amino}-5-phenylpyrimidin-4-yl)benzyl]-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine;1-[4-(2-{[2-(dimethylammonio)ethyl]amino}-5-phenylpyrimidin-4-yl)benzyl]-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine;and1-(4-{2-[[3-(dimethylammonio)propyl](methyl)amino]-5-phenylpyrimidin-4-yl}benzyl)-4-(5-pyridin-2-yl-4h-1,2,4-triazol-3-yl)piperidine;or a stereoisomer thereof.
 7. A pharmaceutical composition comprising apharmaceutical carrier, and dispersed therein, a therapeuticallyeffective amount of a compound of claim
 1. 8. (canceled)
 9. Apharmaceutical composition made by combining the compound of claim 1 anda pharmaceutically acceptable carrier.